Methods for treating IL-18 mediated disorders

ABSTRACT

The invention pertains to methods for treating medical disorders characterized by elevated levels or abnormal expression of IL-18 by administering an IL-18 antagonist, such as soluble IL-18 receptor, a soluble IL-18 binding protein and/or an antibody.

[0001] This application claims the benefit of U.S. provisionalapplication No. 60/241,408, filed Oct. 18, 2000.

FIELD OF THE INVENTION

[0002] The invention pertains to methods for treating certain diseasesand disorders associated with inflammatory and immunoregulatoryresponses. More particularly, the present invention involves treatingdiseases characterized by IL-18 production by administering an IL-18antagonist to an individual afflicted with such a disease.

BACKGROUND

[0003] IL-18, a cytokine produced by activated macrophages and othercells, induces natural killer cell cytotoxicity and participates in thepolarization of the T-lymphocyte helper type 1 phenotype. In addition,IL-18 induces interferon-γ (IFNγ) production in antigen-stimulatedT-cell lines, and acts synergistically with IL-12 to stimulate IFNγproduction in Th1 clones.

[0004] Elevated levels of IL-18 have been observed in various diseasestates including Crohn's disease and rheumatoid arthritis (RA). Forexample, Pallone and co-workers found that IL-18 was upregulated inmucosal intestinal tissue and lamina propria mononuclear cells frompatients with Crohn's disease as compared to samples from patientswithout an inflammatory bowel disease (Monteleone et al., 1999, J.Immunol. 63:143-7). Another group which reported similar results intheir studies of Crohn's disease specimens, also reported a trend ofincreased IL-18 expression in colonic surgical specimens from patientswith ulcerative colitis (Pizzarro et al., 1999, J. Immunol.162:6829-35). In studies of RA, McInnes and coworkers reported thatIL-18 was expressed in RA synovial membrane, and that administration ofrecombinant IL-18 can promote erosive collagen-induced arthritis in ananimal model (when administered immediately before and during collagenpriming and challenge) (Leung et al., 1999, J. Immunol. 164:6495-6502;Gracie et al., 1999, J. Clin. Invest. 104:1393-1401).

[0005] These results led the above-cited authors to speculate that IL-18may play a key pathogenic role in initiating such Th1-mediateddisorders. However, another investigator warned that, because of thepleiotrophic roles that IL-18 is likely to play, one should not concludethat blocking IL-18 would help in treating, for example, rheumatoidarthritis (Dayer, 1999, J. Clin. Invest. 104:1337-1339). In addition toits roles in host defense and the suppression of allergies, IL-i 8induces nitric oxide (NO) production. Dayer reasoned that induction ofNO may be a counter-regulatory loop for IL-18 because NO inhibits theIL-1β-converting enzyme ICE, and thus blocks the processing of proIL-18into a biologically active cytokine (Id.). By inducing NO, IL-18decreases its own activity (Id.). Therefore, inhibition of IL-18 couldincrease ICE activity and promote the maturation of IL-18 and IL-1 β,thereby promoting inflammation and tissue destruction (Id.).

[0006] Thus, the art showed that it was unclear whether attempts todecrease IL-18 would actually lead to therapeutic results in diseasessuch as arthritis and inflammatory bowel diseases. Accordingly, there isa need in the art to resolve this dilemma.

SUMMARY OF THE INVENTION

[0007] The invention is based, in part, on the discovery through actualin vivo experimentation that inhibition of IL-18 can indeed be used totreat inflammatory diseases. Therefore, provided herein are methods fortreating medical disorders associated with IL-18 mediated inflammatoryreactions and/or IL-18 mediated immunoregulatory reactions. The methodsof the present invention include administering an IL-18 antagonist thatinhibits IL-18 inflammatory and/or immunoregulatory signaling to anindividual afflicted with an inflammatory and/or immunoregulatorydisease mediated by IL-18. More particularly, the present inventioninvolves administering an IL-18 antagonist such as, for example, asoluble IL-18 receptor, an IL-18 binding protein, and/or an antibody, tosuch an individual, for a period of time sufficient to induce asustained improvement in the patient's condition. The invention alsoprovides, in part, the use of an IL-18 antagonist in the manufacture ofa medicament for the treatment of medical disorders associated withIL-18 mediated inflammatory reactions and/or IL-18 mediatedimmunoregulatory reactions.

BRIEF DESCRIPTION OF THE FIGURES

[0008]FIG. 1. Effect of IL-18BP-Fc and M147 Administration on WeightLoss in Mouse Model of Inflammatory Bowel Disease. This figure is agraph of the average % weight change of mice in each treatment group(n=8) as a function of days of treatment with 2% DSS, or no DSS, in thedrinking water. Treatment groups were as follows: no DSS, filledsquares; 2% DSS+Human IgG control antibody (250 μg/day), open squares;2% DSS+IL-18BP-Fc fusion protein (600 μg/day), triangles; 2% DSS+M147antibody (250 μg/day), circles.

[0009]FIG. 2. Cytokine Production by MLN Cells After Stimulation WithCD3. FIG. 2A illustrates the average level (n=4) of IFNγ production byMLN cells from each treatment group after stimulation by CD3. FIG. 2Billustrates the average level (n=4) of IL-10 production by MLN cellsfrom each treatment group after stimulation by CD3. Treatment groups areindicated and were: no DSS; 2% DSS +Human IgG control antibody (250μg/day); 2% DSS+IL-18BP-Fc fusion protein (600 μg/day); 2% DSS+M147antibody (250 μg/day).

[0010]FIG. 3. RNase Protection Assays (RPA) Of mRNA Isolated From LargeIntestine. The relative levels of mRNAs in the large intestine, asmeasured by RPA, encoding for IFNγ, TNFα, IL-6, IL-10, IL-18 and IL-1RAare shown in FIG. 3A. The relative levels of mRNAs in the largeintestine encoding for IL-1α and IL-1β are shown in FIG. 3B. Treatmentgroups were as follows: no DSS, purple bars; 2% DSS+Human IgG controlantibody (250 μg/day), black bars; 2% DSS+IL-18BP-Fc fusion protein (600μg/day), blue bars; 2% DSS+M147 antibody (250 μg/day), orange bars.

DETAILED DESCRIPTION OF THE INVENTION

[0011] The present invention provides methods for treating an individualincluding a human, who is suffering from a medical disorder associatedwith IL-18 mediated inflammatory reactions or IL-18 mediatedimmunoregulatory reactions. For purposes of this disclosure, the terms“illness,” “disease,” “medical condition” or “abnormal condition” areused interchangeably with the term “medical disorder.”

[0012] Basis, in part, for the invention is the discovery thatinhibitors of IL-18 are effective in vivo for treating diseases.Specifically, an IL-18 antagonist fusion protein, IL-18BP-Fc, was foundto be useful in preventing experimentally-induced rheumatoid arthritisin a mouse model of this disease. Moreover, the IL-18 antagonist alsoinhibited the progression of an already established disease in the sameanimal model. In addition, two different IL-18 antagonists, a viral p13protein and an IL-18BP-Fc, were also found to be beneficial inameliorating the deleterious effects of two different animal models ofinflammatory bowel diseases. Thus, these in vivo data indicate thatinhibition of IL-18 is effective for treating arthritis, rheumaticdiseases, and inflammatory gastrointestinal diseases. Any method thatneutralizes IL-18 activity or inhibits expression of the IL-18 gene(either transcription or translation) can be used to reduce theinflammatory response caused by IL-18.

[0013] The subject methods involve administering to the patient an IL-18antagonist that is capable of reducing the effective amount ofendogenous biologically active IL-18, such as by reducing the amount ofIL-18 produced, or by preventing the binding of IL-18 to its cellsurface receptor. Such antagonists include receptor-binding peptidefragments of IL-18, IL-18 binding proteins, antibodies directed againstIL-18 or a subunit of the IL-18 receptor, inhibitors (e.g., smallmolecules and peptides) of IL-18 receptor aggregation and signaltransduction, and recombinant proteins comprising all or portions of areceptor for IL-18 or modified variants thereof, includinggenetically-modified muteins, multimeric forms and sustained-releaseformulations. Particular antagonists include IL-18 binding protein,antagonistic IL-18 receptor antibodies and soluble forms of an IL-18receptor. Further, suitable IL-18 antagonists encompass chimericproteins that include portions of both an antibody molecule and an IL-18antagonist molecule. Such chimeric molecules may form monomers, dimersor higher order multimers. Other suitable IL-18 antagonists includepeptides derived from IL-18 that are capable of binding competitively tothe IL-18 signaling receptor, yet do not induce signaling, and nucleicacid based antagonists.

[0014] In a preferred aspect, protein-based therapeutics can be used toinhibit the activity of IL-18 protein. For example, preferred methods ofthe invention utilize IL-18 receptor in a form that binds IL-18, andblocks IL-18 signal transduction, thereby interrupting theproinflammatory and immunoregulatory effects of IL18. PCT Publication WO99/37772, incorporated in its entirety by reference herein, describesthe IL-18 receptor, which is a heterodimeric protein containing an IL-18binding subunit termed IL-1Rrp1, and an accessory subunit termed AcPL.Although the IL-Rrp1 subunit alone will bind IL-18, its affinity forIL-18 is increased dramatically when present in a heterodimeric complexwith the AcPL subunit.

[0015] The IL-1Rrp1 polynucleotide sequence and the amino acid sequencethat it encodes are provided herein as SEQ ID NO: 3 and SEQ ID NO: 4,respectively. The soluble extracellular portion of the IL-1Rrp 1 subunitthat binds IL-18 is represented by amino acids 20 to 329 of SEQ ID NO:4; cleavage of the signal sequence occurs just after amino acid residue19 of SEQ ID NO: 4. However, fragments as small as amino acid residues20 to 123 and amino acid residues 20 to 226 of SEQ ID NO: 4 have beenreported to bind IL-18 and can also be used. The IL-1Rrp1 polypeptide isalso described in U.S. Pat. No. 5,776,731, incorporated in its entiretyby reference herein.

[0016] The AcPL polynucleotide sequence and the amino acid sequence thatit encodes are provided herein as SEQ ID NO: 1 and SEQ ID NO: 2,respectively. The mature extracellular domain of AcPL consists of aminoacids 15 to 356 of SEQ ID NO: 2; cleavage of the signal sequence occursjust after amino acid residue 14 of SEQ ID NO: 2. The AcPL polypeptide,and soluble extracellular fragments thereof, are also described in WO99/37773, incorporated in its entirety by reference herein. Preferableforms of the IL-18 receptor polypeptides are truncated soluble fragmentsthat retain the capability of binding IL-18. Soluble IL-18 receptormolecules include, for example, analogs or fragments of native IL-18receptor having at least 20 amino acids, preferably at least 100 aminoacids, that lack the transmembrane regions of the native molecule, andthat are capable of binding IL-18.

[0017] One preferred soluble form of an IL-18 receptor for use in themethods of the present invention includes amino acids 1-329 (20-329after cleavage of the signal sequence) of SEQ ID NO: 4. An even morepreferred soluble form of IL-18 receptor is a heterodimeric receptorthat includes at least amino acid residues 20-123, 20-226 or 20-329 ofSEQ ID NO: 4 (the IL-1Rrp1 subunit), and at least amino acids 15-340 ofSEQ ID NO: 2 (the AcPL subunit), in a covalent or non-covalentassociation.

[0018] Another preferred soluble IL-18 antagonist for use in the methodsof the present invention is the IL-18 binding protein. PCT PublicationWO 99/09063 describes the IL-18 binding protein, including usefulsoluble fragments thereof, and this description is incorporated byreference herein. A particularly useful form of the IL-18 bindingprotein is a fusion with an Fc domain of an antibody. The amino acidsequence of an example of such a fusion protein, termed IL-18BP-Fcherein, is presented in SEQ ID NO: 5. This 422 amino acid protein, whenexpressed in a mammalian cell, will be secreted; the mature secretedform of the protein contains amino acid residues 29-422. Of theseresidues, amino acid residues 29-192 represent the IL-18 binding proteinportion of the molecule, and amino acid residues 193-422 represent theFc portion of the molecule. The Fc region facilitates purification anddimerization of the fusion polypeptide.

[0019] Antagonists derived from IL-18 receptors and IL-18 bindingprotein (e.g. soluble forms that bind IL-18) compete for IL-18 withIL-18 receptors on the cell surface, thus inhibiting IL-18 from bindingto cells, thereby preventing it from manifesting its biologicalactivities. Binding of soluble IL-18 receptor or IL-18 binding proteincan be assayed using ELISA or any other convenient assay.

[0020] Other types of protein-based therapeutics are antibodies thatspecifically recognize one or more epitopes of IL-18, or epitopes ofconserved variants of IL-18, or peptide fragments of the IL-18polypeptide that competitively inhibit IL-18 activity. Antibodies toIL-18 can most conveniently be raised to a recombinantly produced formof the protein. For example, human IL-18 has been recombinantly producedfrom both a cloned cDNA (Ushio et al., 1996, J. Immunol. 156:4274-4279)and cloned genomic DNA (U.S. Pat. No. 6,060,283). Or, antibodies thatspecifically recognize a component of the IL-18 receptor and thatprevent signaling through the receptor by IL-18 can be used to inhibitIL-18 activity. IL-18 antagonists that are antibodies include but arenot limited to polyclonal antibodies, monoclonal antibodies (mAbs),humanized or chimeric antibodies, single chain antibodies, Fabfragments, F(ab′)₂ fragments, fragments produced by a Fab expressionlibrary, anti-idiotypic (anti-Id) antibodies, and epitope-bindingfragments of any of the above. Thus, such antibodies can, therefore, beutilized as part of inflammatory disorder treatment methods.

[0021] For the production of antibodies, various host animals can beimmunized by injection with the IL-18 polypeptide, truncated IL-18polypeptides, a component of the IL-18 receptor (e.g., the IL-18 bindingsubunit, or the AcPL subunit), a truncated version of a component of theIL-18 receptor, and functional equivalents and mutants thereof. Suchhost animals may include but are not limited to rabbits, mice, and rats,to name but a few. Various adjuvants may be used to increase theimmunological response, depending on the host species, including but notlimited to Freund's (complete and incomplete), mineral gels such asaluminum hydroxide, surface active substances such as lysolecithin,pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpethemocyanin, dinitrophenol, and potentially useful human adjuvants suchas BCG (bacille Calmette-Guerin) and Corynebacterium parvum.Alternatively, libraries of antibody fragments can be screened and usedto develop human antibodies through recombinant techniques. Suchlibraries are commercially available from, for example, CambridgeAntibody Technology (Melbourne, UK), and Morphosys (Munich, Del.).

[0022] Monoclonal antibodies, which are homogeneous populations ofantibodies to a particular antigen, can be obtained by any techniquethat provides for the production of antibody molecules by continuouscell lines in culture. These include, but are not limited to, thehybridoma technique of Kohler and Milstein, (U.S. Pat. No. 4,376,110),the human B-cell hybridoma technique (Kosbor et al., 1983, ImmunologyToday 4:72; Cole et al., 1983, Proc. Natl. Acad. Sci. USA 80:2026-2030),and the EBV-hybridoma technique (Cole et al., 1985, MonoclonalAntibodies And Cancer Therapy, Alan R. Liss, Inc., pp. 77-96). Suchantibodies may be of any immunoglobulin class including IgG, IgM, IgE,IgA, IgD and any subclass thereof. The hybridoma producing the mAb maybe cultivated in vitro or in vivo. Or, the antibody genes can be clonedand optionally otherwise altered, and expressed in another cell lineapproved for recombinant production of protein pharmaceuticals such as,for example, CHO cells.

[0023] In addition, techniques developed for the production of “chimericantibodies” (Takeda et al., 1985, Nature, 314:452-454) by splicing thegenes from a mouse antibody molecule of appropriate antigen specificitytogether with genes from a human antibody molecule of appropriatebiological activity can be used. A chimeric antibody is a molecule inwhich different portions are derived from different animal species, suchas those having a variable region derived from a porcine mAb and a humanimmunoglobulin constant region.

[0024] Preferably, for use in humans, the antibodies are human orhumanized; techniques for creating such human or humanized antibodiesare also well known and are commercially available from, for example,Protein Design Labs, Inc. (Fremont, Calif.), Medarex Inc. (Princeton,N.J.) and Abgennix Inc. (Fremont, Calif.).

[0025] Techniques described for the production of single chainantibodies (U.S. Pat. No. 4,946,778; Bird, 1988, Science 242:423-426;Huston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883; and Wardet al., 1989, Nature 334:544-546) can also be adapted to produce singlechain antibodies against IL-18 gene products. Single chain antibodiesare formed by linking the heavy and light chain fragments of the Fvregion via an amino acid bridge, resulting in a single chainpolypeptide.

[0026] Antibody fragments which recognize specific epitopes can begenerated by known techniques. For example, such fragments include butare not limited to: the F(ab′)₂ fragments which can be produced bypepsin digestion of the antibody molecule and the Fab fragments whichcan be generated by reducing the disulfide bridges of the (ab′)₂fragments. Alternatively, Fab expression libraries can be constructed(Huse et al., 1989, Science, 246:1275-1281) to allow rapid and easyidentification of monoclonal Fab fragments with the desired specificity.

[0027] Still another IL-18 antagonist that can be used in the methodsand compositions of the invention is a virally encoded IL-18 bindingprotein. For example, the fowlpox (ectromelia) virus p13 polypeptide hasbeen shown to bind to, and inhibit the biological activity, of IL-18(Born et al., 2000, J. Immunol. 164(6):3246-54, incorporated in itsentirety by reference herein). The invention also encompasses the use ofIL-18 antagonists yet to be discovered in the therapeutic methods andcompositions.

[0028] In alternative embodiments, nucleic acid-based immuno therapy canbe designed to reduce the level of endogenous IL-18 gene expression,e.g., using antisense or ribozyme approaches to inhibit or preventtranslation of IL-18 mRNA transcripts; triple helix approaches toinhibit transcription of the IL-18 gene; or targeted homologousrecombination to inactivate or “knock out” the IL-18 gene or itsendogenous promoter.

[0029] Antisense approaches involve the design of oligonucleotides(either DNA or RNA) that are complementary to IL-18 mRNA. The antisenseoligonucleotides will bind to the complementary IL-18 mnRNA transcriptsand prevent translation. The IL-18 cDNA sequence is described in Ushioet al., 1996, J. Immunol. 156:42744279.

[0030] Absolute complementarity to the mRNA transcript, althoughpreferred, is not required. A sequence “complementary” to a portion ofan RNA, as referred to herein, means a sequence having sufficientcomplementarity to be able to hybridize with the RNA, forming a stableduplex. In the case of double-stranded antisense nucleic acids, a singlestrand of the duplex DNA can thus be tested, or triplex formation can beassayed. The ability to hybridize will depend on both the degree ofcomplementarity and the length of the antisense nucleic acid.

[0031] Oligonucleotides that are complementary to the 5′ end of themessage, e.g., the 5′ untranslated sequence up to and including the AUGinitiation codon, should work most efficiently at inhibitingtranslation. However, oligonucleotides complementary to either the 5′-or 3′-non-translated, and any of the coding and/or non-coding regions ofthe IL-18 gene transcript could be used in an antisense approach toinhibit translation of endogenous IL-18 mRNA. Antisense nucleic acidsshould be at least six nucleotides in length, and are preferablyoligonucleotides ranging from 6 to about 50 nucleotides in length. Inspecific aspects the oligonucleotide is at least 10 nucleotides, atleast 17 nucleotides, at least 25 nucleotides or at least 50nucleotides.

[0032] The oligonucleotides can be DNA or RNA or chimeric mixtures orderivatives or modified versions thereof, single-stranded ordouble-stranded. The oligonucleotide can be modified at the base moiety,sugar moiety, or phosphate backbone, for example, to improve stabilityof the molecule, hybridization, etc. The oligonucleotide can includeother appended groups such as peptides (e.g., for targeting host cellreceptors in vivo), or agents facilitating transport across the cellmembrane such as lipid carriers (see, e.g., Letsinger et al., 1989,Proc. Natl. Acad. Sci. U.S.A. 86:6553-6556; Lemaitre et al., 1987, Proc.Natl. Acad. Sci. 84:648-652; PCT Publication No. WO88/09810, publishedDec. 15, 1988), or hybridization-triggered cleavage agents orintercalating agents. (See, e.g., Zon, 1988, Pharm. Res. 5:539-549).

[0033] Oligonucleotides can be synthesized by standard methods known inthe art, e.g. by use of an automated DNA synthesizer (such as arecommercially available from Biosearch, Applied Biosystems, etc.). Asexamples, phosphorothioate oligonucleotides can be synthesized by themethod of Stein et al., 1988, Nucl. Acids Res. 16:3209.Methylphosphonate oligonucleotides can be prepared by use of controlledpore glass polymer supports (Sarin et al., 1988, Proc. Natl. Acad. Sci.U.S.A. 85:7448-7451).

[0034] The antisense molecules should be delivered to cells that expressthe IL-18 transcript in vivo. A number of methods have been developedfor delivering antisense DNA or RNA to cells; e.g., antisense moleculescan be injected directly into the tissue or cell derivation site, ormodified antisense molecules, designed to target the desired cells(e.g., antisense linked to peptides or antibodies that specifically bindreceptors or antigens expressed on the target cell surface) can beadministered systemically.

[0035] However, it is often difficult to achieve intracellularconcentrations of the antisense sufficient to suppress translation ofendogenous mRNAs. Therefore a preferred approach utilizes a recombinantDNA construct in which the antisense sequence is placed under thecontrol of a strong pol III or pol II promoter. The use of such aconstruct to transfect target cells in the patient will result in thetranscription of sufficient amounts of single stranded RNAs that willform complementary base pairs with the endogenous IL-18 gene transcriptsand thereby prevent translation of the IL-18 mRNA. For example, a vectorcan be introduced in vivo such that it is taken up by a cell and directsthe transcription of an antisense RNA. Such a vector can remain episomalor become chromosomally integrated, as long as it can be transcribed toproduce the desired antisense RNA. Such vectors can be constructed byrecombinant DNA technology methods standard in the art. Vectors can beplasmid, viral, or others known in the art, used for replication andexpression in mammalian cells.

[0036] Ribozyme molecules designed to catalytically cleave IL-18 mRNAtranscripts can also be used to prevent translation of IL-18 mRNA andexpression of IL-18 protein. (See, e.g., PCT International PublicationWO90/11364; U.S. Pat. No. 5,824,519). The ribozymes that can be used inthe present invention include hammerhead ribozymes (Haseloff andGerlach, 1988, Nature, 334:585-591), RNA endoribonucleases (hereinafter“Cech-type ribozymes”) such as the one which occurs naturally inTetrahymena thermophila (known as the IVS, or L-19 IVS RNA) and whichhas been extensively described by Thomas Cech and collaborators(International Patent Application No. WO 88/04300; Been and Cech, 1986,Cell 47:207-216).

[0037] As in the antisense approach, the ribozymes can be composed ofmodified nucleotides (e.g. for improved stability, targeting, etc.) andshould be delivered to cells which express the IL-18 polypeptide invivo. A preferred method of delivery involves using a DNA construct“encoding” the ribozyme under the control of a strong constitutive polIII or pol II promoter, so that transfected cells will producesufficient quantities of the ribozyme to destroy endogenous IL-18polypeptide messages and inhibit translation. Because ribozymes, unlikeantisense molecules, are catalytic, a lower intracellular concentrationis required for efficiency.

[0038] This invention additionally provides for the use of IL-18antagonists in the manufacture of a medicament for the treatment ofnumerous diseases. This invention additionally provides for the use ofpolynucleotides encoding IL-18 antagonists in the manufacture of IL-18antagonists for use in the manufacture of a medicament for the treatmentof diseases disclosed herein.

[0039] Soluble IL-18 antagonists that are polypeptides suitable in thepractice of this invention can be fused with a second polypeptide toform a chimeric protein. In one embodiment of such a chimeric protein,the second polypeptide can promote the spontaneous formation by thechimeric protein of a dimer, trimer or higher order multimer that iscapable of binding IL-18 molecule and preventing it from binding to acell-bound receptor that promotes IL-18 signaling. For example, chimericproteins used as antagonists can be proteins that contain portions ofboth an antibody molecule and a soluble IL-18 antagonist. In particularaspects, the Fc portion of an antibody molecule can be used. Onesuitable Fc polypeptide, described in PCT application WO 93/10151(hereby incorporated by reference), is a single chain polypeptideextending from the N-terminal hinge region to the native C-terminus ofthe Fc region of a human IgG1 antibody. Another useful Fc polypeptide isthe Fc mutein described in U.S. Pat. No. 5,457,035 and in Baum et al.,1994, EMBO J. 13:3992-4001. Another example of a oligomerization domainis a leucine zipper, the use of which is well known in the art. Indeed,any oligomerization domain known or yet to be discovered can be used asthe second polypeptide. One preferred oligomer IL-18 antagonist suitablefor treating diseases in humans and other mammals is the IL-18BP-Fc(IL-18 binding protein fused to an Fc mutein region; SEQ ID NO: 5) whoseuse is illustrating below by way of non-limiting working examples. Otherpreferred oligomer IL-18 antagonists are any of the soluble IL-18receptor molecules described above fused to either an Fc mutein regionor a leuzine zipper or any other oligomerization domain.

[0040] In one preferred embodiment of the invention, sustained-releaseforms of soluble IL-18 antagonists, and in particular, soluble IL-18receptor or IL-18 binding protein, are used. Sustained-release formssuitable for use in the disclosed methods include, but are not limitedto, IL-18 antagonists that are encapsulated in a slowly-dissolvingbiocompatible polymer, admixed with such a polymer, and or encased in abiocompatible semi-permeable implant. In addition, the IL-18 antagonistcan be conjugated with polyethylene glycol (pegylated) to prolong itsserum half-life or to enhance protein delivery.

[0041] To treat a medical disorder characterized by abnormal or excessexpression of IL-18 or abnormal or excess IL-18 signaling, a moleculecomprising an IL-18 antagonist, preferably a soluble IL-18 receptor orIL-18 binding protein, or an antibody, is administered to the patient inan amount and for a time sufficient to induce a sustained improvement inat least one indicator that reflects the severity of the disorder. Animprovement is considered “sustained” if the patient exhibits theimprovement on at least two occasions separated by one to four weeks.The degree of improvement is determined based on signs or symptoms, andmay also employ questionnaires that are administered to the patient,such as quality-of-life questionnaires. A therapeutically effectiveamount of an IL-18 antagonist is that sufficient to achieve such asustained improvement.

[0042] Various indicators that reflect the extent of the patient'sillness may be assessed for determining whether the amount and time ofthe treatment is sufficient. The baseline value for the chosen indicatoror indicators is established by examination of the patient prior toadministration of the first dose of the soluble IL-18 receptor or otherIL-18 antagonist. Preferably, the baseline examination is done withinabout 60 days of administering the first dose.

[0043] Improvement is induced by repeatedly administering a dose ofIL-18 antagonist until the patient manifests an improvement overbaseline for the chosen indicator or indicators. In treating chronicconditions, this degree of improvement is obtained by repeatedlyadministering this medicament over a period of at least a month or more,e.g., for one, two, or three months or longer, or indefinitely. A periodof one to six weeks, or even a single dose, often is sufficient fortreating acute conditions.

[0044] Although the extent of the patient's illness after treatment mayappear improved according to one or more indicators, treatment may becontinued indefinitely at the same level or at a reduced dose orfrequency. Once treatment has been reduced or discontinued, it later maybe resumed at the original level if symptoms should reappear.

[0045] Any efficacious route of administration can be used totherapeutically administer a soluble IL-18 receptor or other IL-18antagonists. If injected, an IL-18 antagonist can be administered, forexample, via intra-articular, intravenous, intramuscular, intralesional,intraperitoneal or subcutaneous routes by bolus injection or bycontinuous infusion. Other suitable means of administration includesustained release from implants, aerosol inhalation, eyedrops, oralpreparations, including pills, syrups, lozenges or chewing gum, topicalpreparations such as lotions, gels, sprays, ointments, buccalpreparations, or other suitable techniques. Alternatively, IL-18antagonist polypeptides, such as a soluble IL-18 receptor or IL-18binding protein, can be administered by implanting cultured cells thatexpress the protein; for example, by implanting cells which express asoluble IL-18 receptor or an IL-18 binding protein. In one embodiment,the patient's own cells are induced to produce by transfection in vivoor ex vivo with a polynucleotide that encodes an IL-18 antagonist, andparticularly soluble IL-18 receptor or IL-18 binding protein. Thispolynucleotide can be introduced into the patient's cells, for example,by injecting naked DNA or liposome-encapsulated DNA that encodes solubleIL-18 receptor or other selected IL-18 antagonist, or by other means oftransfection. When an IL-18 antagonist is administered in combinationwith one or more other biologically active compounds, these can beadministered by the same or by different routes, and can be administeredsimultaneously, separately or sequentially.

[0046] Soluble IL-18 receptor or IL-18 binding protein or otherantagonists of IL-18 preferably are administered in the form of aphysiologically acceptable composition comprising purified recombinantprotein in conjunction with physiologically acceptable carriers,excipients or diluents. Such carriers are nontoxic to recipients at thedosages and concentrations employed. Ordinarily, preparing suchcompositions entails combining the IL-18 antagonist with buffers,antioxidants such as ascorbic acid, low molecular weight polypeptides(such as those having fewer than 10 amino acids), proteins, amino acids,carbohydrates such as glucose, sucrose or dextrins, chelating agentssuch as EDTA, glutathione and other stabilizers and excipients. Neutralbuffered saline or saline mixed with conspecific serum albumin areexemplary appropriate diluents. The IL-18 antagonist preferably isformulated as a lyophilizate using appropriate excipient solutions(e.g., sucrose) as diluents. Appropriate dosages can be determined instandard dosing trials, and may vary according to the chosen route ofadministration. In accordance with appropriate industry standards,preservatives may also be added, such as benzyl alcohol. The amount andfrequency of administration will depend, of course, on such factors asthe nature and severity of the indication being treated, the desiredresponse, the age and condition of the patient, and so forth.

[0047] In one embodiment of the invention, IL-18 antagonist isadministered one time per week to treat the various medical disordersdisclosed herein, in another embodiment is administered at least twotimes per week, and in another embodiment is administered at least onceper day. An adult patient is a person who is 18 years of age or older.If injected, the effective amount, per adult dose, of an IL-18 bindingprotein or an IL-18 receptor protein ranges from 1-200 mg/m², or from1-40 mg/m² or about 5-25 mg/m². Alternatively, a flat dose may beadministered, whose amount may range from 2-400 mg/dose, 2-100 mg/doseor from about 10-80 mg/dose. If the dose is to be administered more thanone time per week an exemplary dose range is the same as the foregoingdescribed dose ranges or lower. Preferably, such IL-18 antagonists areadministered two or more times per week at a per dose range of 25-100mg/dose. In one embodiment of the invention, the various indicationsdescribed below are treated by administering a preparation acceptablefor injection containing an IL-18 binding protein at 80-100 mg/dose, oralternatively, containing 80 mg per dose. If the IL-18 antagonist is anantibody, the dose can be from 0.1 to 10 mg/kg, preferably givenintravenously as a 15 minute to 3 hour infusion. The dose isadministered repeatedly at biweekly, weekly, or separated by several(2-8 weeks).

[0048] If a route of administration of IL-18 antagonist other thaninjection is used, the dose is appropriately adjusted in accord withstandard medical practices. For example, if the route of administrationis inhalation, dosing may be one to seven times per week at dose rangesfrom 10 mg/dose to 50 mg per dose.

[0049] In many instances, an improvement in a patient's condition willbe obtained by injecting a dose of up to about 100 mg of a soluble IL-18receptor or IL-18 binding protein or an antagonistic antibody one tothree times per week over a period of at least three weeks, thoughtreatment for longer periods may be necessary to induce the desireddegree of improvement. For incurable chronic conditions, the regimen maybe continued indefinitely.

[0050] For pediatric patients (age 4-17), a suitable regimen involvesthe subcutaneous injection of 0.4 mg/kg to 5 mg/kg of IL-18 receptor orIL-18 binding protein, administered by subcutaneous injection one ormore times per week.

[0051] The invention further includes the administration of IL-18antagonist concurrently with one or more other drugs that areadministered to the same patient, each drug being administered accordingto a regimen suitable for that medicament. This encompassespre-treatment, simultaneous treatment, sequential treatment andalternating regimens. Examples of such drugs include but are not limitedto antivirals, antibiotics, analgesics, corticosteroids, antagonists ofinflammatory cytokines, DMARDs and non-steroidal anti-inflammatories.Additionally, one type of IL-18 antagonist can be combined with a secondIL-18 antagonist, including an antibody against IL-18 or against anIL-18 receptor, additional IL-18 receptor derivatives, or othermolecules that reduce endogenous IL-18 levels, such as peptidomimeticIL-18 antagonists.

[0052] In one preferred embodiment of the invention, the various medicaldisorders disclosed herein as being treatable with an IL-18 antagonistare treated in combination with another cytokine or cytokine inhibitor.For example, IL-18 antagonist can be administered in a composition thatalso contains a compound that inhibits the interaction of otherinflammatory cytokines with their receptors. The IL-18 antagonist andother cytokine inhibitor can be administered as separate compositions,and these can be administered by the same or different routes. Examplesof cytokine inhibitors used in combination with IL-18 antagonist includethose that antagonize, for example, IFNγ, IL-6, IL-8, IL-12, IL-15 andTNF, particularly TNFα. Anti-inflamatory cytokines include but are notlimited to IL-4, TGFβ, and EGF. Other combinations for treating thehereindescribed diseases include the use of IL-18 antagonist withcompounds that interfere with the binding of RANK and RANK-ligand, suchas RANK-ligand inhibitors, or soluble forms of RANK, including RANK:Fc.For example, the combination of IL-18 antagonist and RANK:Fc are usefulfor preventing bone destruction in various settings including but notlimited to various rheumatic disorders. Still another combination fortreating the hereindescribed diseases include the use of an IL-18antagonist in combination with an IL-1 antagonist, such as, for example,a soluble IL-1 receptor type II molecule or an antagonistic antibody tothe IL-1 receptor. In addition, IL-18 antagonist can be administered incombination with soluble forms of an IL-17 receptor (such as IL-17R:Fc),IL-12 binding protein, or antibodies against CD30-ligand or against CD4.

[0053] The present invention further encompasses methods for treatingthe herein disclosed medical disorders with a combination of an IL-18antagonist and a TNF inhibitor, preferably TNFR:Fc (ENBREL® marketed byImmunex Corp) and optionally with any combination of the above describedcytokines or cytokine inhibitors that are active agents in combinationtherapies. For example, in accordance with the present invention,combination therapy methods for treating rheumatic, arthritic andvarious inflammatory gastrointestinal diseases include administeringIL-18 antagonist and ENBREL®. Thus, the present invention also relatesto the using IL-18 antagonists and TNF inhibitors in combinationtherapies for use in medicine and in particular in therapeutic andpreventive therapies for the medical disorders described herein. The usein medicine can involve the treatment of any of the medical disorders asdescribed herein with a combination therapy that includes administeringa combination of an IL-18 antagonist and ENBREL®. The IL-18 antagonists(e.g., soluble IL-18 receptor or IL-18 binding protein or anantagonistic antibody) and TNF inhibitor (e.g., ZENBREL® may be in theform of compounds, compositions or combination therapies. Where thecompounds are used together with one or more other components, thecompound and the one or more other components can be administeredsimultaneously, separately or sequentially (usually in pharmaceuticalformat).

[0054] The present invention also relates to the use of IL-18antagonists (as disclosed), such as, for example, a soluble IL-18receptor, in the manufacture of a medicament for the prevention ortherapeutic treatment of each medical disorder disclosed herein.

[0055] Conditions of the gastrointestinal system are treatable orpreventable with IL-18 antagonists, compositions or combinationtherapies, including coeliac disease. For example, IL-18 antagonistcompositions, with or without TNF inhibitors (ENBREL®) or other activeagents described above are suitable for treating or preventing coeliacdisease. In addition, the compounds, compositions and combinationtherapies of the invention are suitable for treating or preventingCrohn's disease; ulcerative colitis; idiopathic gastroparesis;pancreatitis, including chronic pancreatitis; inflammatory bowel diseaseand ulcers, including gastric and duodenal ulcers.

[0056] Other embodiments of the present invention include methods forusing the disclosed IL-18 antagonists, in particular soluble IL-18receptor, compositions or combination therapies, e.g. soluble IL-18receptor and ENBREL®, to treat or prevent a variety of rheumaticdisorders. These include adult and juvenile rheumatoid arthritis;scleroderma; systemic lupus erythematosus; gout; osteoarthritis;polymyalgia rheumatica; seronegative spondylarthropathies, includingankylosing spondylitis, and Reiter's disease. The subject IL-18antagonists, compositions and combination therapies are used also totreat psoriatic arthritis and chronic Lyme arthritis. Also treatable orpreventable with these compounds, compositions and combination therapiesare Still's disease and uveitis associated with rheumatoid arthritis. Inaddition, the compounds, compositions and combination therapies of theinvention are used in treating disorders resulting in inflammation ofthe voluntary muscle and other muscles, including dermatomyositis,inclusion body myositis, polymyositis, and lymphangioleimyomatosis.

[0057] IL-18 antagonist can be used to treat psoriatic arthritis incombination with one, two, three or more other medications that areeffective against psoriasis. These additional medications can beadministered before, simultaneously with, or sequentially with thesoluble IL-18 receptor. Drugs suitable for combination therapies includepain medications (analgesics), including but not limited toacetaminophen, codeine, propoxyphene napsylate, oxycodone hydrochloride,hydrocodone bitartrate and tramadol. In addition, IL-18 antagonist canbe administered in combination with a soluble TNF receptor (ENBREL®,methotrexate, sulfasalazine, gold salts, azathioprine, cyclosporine,antimalarials, oral steroids (e.g., prednisone) or colchicine.Non-steroidal anti-inflammatories may also be coadministered with theIL-18 antagonist, including but not limited to: salicylic acid(aspirin); ibuprofen; indomethacin; celecoxib; rofecoxib; ketorolac;nambumetone; piroxicam; naproxen; oxaprozin; sulindac; ketoprofen;diclofenac; other COX-1 and/or COX-2 inhibitors, salicylic acidderivatives, propionic acid derivatives, acetic acid derivatives,fumaric acid derivatives, carboxylic acid derivatives, butyric acidderivatives, oxicams, pyrazoles and pyrazolones, including newlydeveloped anti-inflammatories.

[0058] Moreover, the IL-18 antagonist can be used to treat psoriaticarthritis in combination with topical steroids, systemic steroids,antagonists of inflammatory cytokines, antibodies against T cell surfaceproteins, anthralin, coal tar, vitamin D3 and its analogs (including1,25-dihydroxy vitamin D3 and calcipotriene), topical retinoids, oralretinoids (including but not limited to etretinate, acitretin andisotretinoin), topical salicylic acid, methotrexate, cyclosporine,hydroxyurea and sulfasalazine. In addition, it can be administered incombination with one or more of the following compounds; minocycline;misoprostol; oral collagen; penicillamine; 6-mercaptopurine; nitrogenmustard; gabapentin; bromocriptine; somatostatin; peptide T; anti-CD4monoclonal antibody; fumaric acid; polyunsaturated ethyl ester lipids;zinc; and other drugs that can be used to treat psoriasis.

[0059] It is understood that the response by individual patients to theaforementioned medications or combination therapies may vary, and themost efficacious combination of drugs for each patient will bedetermined by his or her physician.

[0060] Further, in addition to human patients, IL-18 antagonists areuseful in the treatment of non-human animals, such as pets (dogs, cats,birds, primates, etc.), domestic farm animals (horses cattle, sheep,pigs, birds, etc.), or any animal that suffers from an IL-18-mediatedinflammatory or arthritic condition. In such instances, an appropriatedose may be determined according to the animal's body weight. Forexample, a dose of 0.2-1 mg/kg may be used. Alternatively, the dose isdetermined according to the animal's surface area, an exemplary doseranging from 0.1-20 mg/m², or more preferably, from 5-12 mg/m². Forsmall animals, such as dogs or cats, a suitable dose is 0.4 mg/kg.Soluble IL-18 receptor (preferably constructed from genes derived fromthe recipient species), or another soluble IL-18 antagonist, isadministered by injection or other suitable route one or more times perweek until the animal's condition is improved, or it may be administeredindefinitely.

[0061] Additional Diseases Treatable By IL-18 Antagonists

[0062] The disclosed experimental data demonstrates that IL-18antagonists can be used to treat inflammatory conditions associated withIL18. Accordingly, a number of other diseases are treatable with IL-18antagonists.

[0063] Cardiovascular disorders are treatable and/or preventable withthe disclosed IL-18 antagonists, pharmaceutical compositions orcombination therapies. In particularly, cardiovascular disorders aretreatable with IL-18 antagonist compositions, alone or in combinationwith TNF inhibitors (e.g. ENBREL) and or other agents as describedabove. Cardiovasuclar disorders thus treatable include aortic aneurysms;arteritis; vascular occlusion, including cerebral artery occlusion;complications of coronary by-pass surgery; ischemia/reperfusion injury;heart disease, including atherosclerotic heart disease, myocarditis,including chronic autoimmune myocarditis and viral myocarditis; heartfailure, including chronic heart failure (CHF), cachexia of heartfailure; myocardial infarction; restenosis and/or atherosclerosis afterheart surgery or after carotid artery balloon angioplastic procedures;silent myocardial ischemia; post implantation complications of leftventricular assist devices; Raynaud's phenomena; thrombophlebitis;vasculitis, including Kawasaki's vasculitis; veno-occlusive disease,giant cell arteritis, Wegener's granulomatosis; mental confusionfollowing cardio pulmonary by pass surgery, and Schoenlein-Henochpurpura. Combinations of IL-18 antagonists, TNF inhibitors andangiogenesis inhibitors (e.g. anti-VEGF) are useful for treating certaincardiovascular diseases such as aortic aneurysms and tumors.

[0064] In addition, the subject IL-18 antagonists, compositions andcombination therapies are used to treat chronic pain conditions, such aschronic pelvic pain, including chronic prostatitis/pelvic pain syndrome.As a further example, soluble IL-18 receptor and the compositions andcombination therapies of the invention are used to treat post-herpeticpain.

[0065] Provided also are methods for using IL-18 antagonists,compositions or combination therapies to treat various disorders of theendocrine system. For example, IL-18 binding protein compositions orother IL-18 antagonist compositions, with or without TNF inhibitors(ENBREL) or other active agents described above, are suitable for use totreat juvenile onset diabetes (includes autoinrnune andinsulin-dependent types of diabetes) and also to treat maturity onsetdiabetes (includes non-insulin dependent and obesity-mediated diabetes).In addition, the subject compounds, compositions and combinationtherapies are used to treat secondary conditions associated withdiabetes, such as diabetic retinopathy, kidney transplant rejection indiabetic patients, obesity-mediated insulin resistance, and renalfailure, which itself may be associated with proteinurea andhypertension. Other endocrine disorders also are treatable with thesecompounds, compositions or combination therapies, including polycysticovarian disease, X-linked adrenoleukodystrophy, hypothyroidism andthyroiditis, including Hashimoto's thyroiditis (i.e., autoimmunethyroiditis). Further, IL-18 antagonists, including IL-18 receptor orIL-18 binding protein, alone or in combination with other cytokines,including TNF inhibitors such as ENBREL, are useful in treating orpreventing medical conditions associated with thyroid cell dysfunction,including euthyroid sick syndrome.

[0066] Included also are methods for using the subject IL-18antagonists, compositions or combination therapies for treatingdisorders of the genitourinary system. For example, IL-18 antagonistcompositions, alone or in combination with TNF inhibitors (ENBREL) orother active agents described above are suitable for treating orpreventing nephrotic syndrome and/or glomerulonephritis, includingautoimmune glomerulonephritis, glomerulonephritis due to exposure totoxins or glomerulonephritis secondary to infections with haemolyticstreptococci or other infectious agents. Also treatable with thecompounds, compositions and combination therapies of the invention areuremic syndrome and its clinical complications (for example, renalfailure, anemia, and hypertrophic cardiomyopathy), including uremicsyndrome associated with exposure to environmental toxins, drugs orother causes and renal failure due to ischemia. IL-18 antagonists,particularly soluble IL-18 receptor or IL-18 binding protein or anantibody, alone or in combination with TNF inhibitors, particularlyENBREL, are useful in treating and preventing complications that arisefrom inflammation of the gallbladder wall that leads to alteration inabsorptive function. Included in such complications are cholelithiasis(gallstones) and choliedocholithiasis (bile duct stones) and therecurrence of cholelithiasis and choliedocholithiasis. Furtherconditions treatable with the compounds, compositions and combinationtherapies of the invention are complications of hemodialysis; prostateconditions, including benign prostatic hypertrophy, nonbacterialprostatitis and chronic prostatitis; and complications of hemodialysis.

[0067] Also provided herein are methods for using IL-18 antagonists,compositions or combination therapies to treat various hematologic andoncologic disorders. For example, soluble IL-18 receptor or IL-18binding protein or an antagonistic antibody, alone or in combinationwith a TNF inhibitor (ENBREL) or other active agents as described above,can be used to treat various forms of cancer, including acutemyelogenous leukemia, chronic myelogenous leukemia leukemia,Epstein-Barr virus-positive nasopharyngeal carcinoma, glioma, colon,stomach, prostate, renal cell, cervical and ovarian cancers, lung cancer(SCLC and NSCLC), including cancer-associated cachexia, fatigue,asthenia, paraneoplastic syndrome of cachexia and hypercalcemia.Additional diseases treatable with the subject IL-18 antagonists,compositions or combination therapies are solid tumors, includingsarcoma, osteosarcoma, and carcinoma, such as adenocarcinoma (forexample, breast cancer) and squamous cell carcinoma. In addition, thesubject compounds, compositions or combination therapies are useful fortreating esophogeal cancer, gastric cancer, leukemia, including acutemyelogenous leukemia, chronic myelogenous leukemia, myeloid leukemia,chronic or acute lymphoblastic leukemia and hairy cell leukemia. Othermalignancies with invasive metastatic potential, including multiplemyeloma, can be treated with the subject compounds, compositions andcombination therapies, and particularly combination therapies thatinclude soluble IL-18 receptor and soluble TNF receptor (ENBREL). Inaddition, the disclosed IL-18 antagonists, compositions and combinationtherapies can be used to treat anemias and hematologic disorders,including anemia of chronic disease, aplastic anemia, includingFanconi's aplastic anemia; idiopathic thrombocytopenic purpura (ITP);thrombotic thrombocytopenic purpura, myelodysplastic syndromes(including refractory anemia, refractory anemia with ringedsideroblasts, refractory anemia with excess blasts, refractory anemiawith excess blasts in transformation); myelofibrosis/myeloid metaplasia;and sickle cell vasocclusive crisis.

[0068] Various lymphoproliferative disorders also are treatable with thedisclosed IL-18 antagonists, compositions or combination therapies.IL-18 antagonist, alone or in combination with a TNF inhibitor, such asENBREL, or other active agents are useful for treating or preventingautoimmune lymphoproliferative syndrome (ALPS), chronic lymphoblasticleukemia, hairy cell leukemia, chronic lymphatic leukemia, peripheralT-cell lymphoma, small lymphocytic lymphoma, mantle cell lymphoma,follicular lymphoma, Burkitt's lymphoma, Epstein-Barr virus-positive Tcell lymphoma, histiocytic lymphoma, Hodgkin's disease, diffuseaggressive lymphoma, acute lymphatic leukemias, T gammalymphoproliferative disease, cutaneous B cell lymphoma, cutaneous T celllymphoma (i.e., mycosis fungoides) and Sézary syndrome.

[0069] In addition, the subject IL-18 antagonists, compositions andcombination therapies are used to treat hereditary conditions. Inparticular, IL-18 antagonist, alone or in combination with a TNFinhibitor such as ENBREL, is useful to treat diseases such as Gaucher'sdisease, Huntington's disease, linear IgA disease, and musculardystrophy.

[0070] Other conditions treatable or preventable by the disclosed IL-18antagonists, compositions and combination therapies include thoseresulting from injuries to the head or spinal cord including subduralhematoma due to trauma to the head. For example, soluble IL-18 receptor,alone or in combination with a TNF inhibitor such as ENBREL are usefulin treating head injuries and spinal chord injuries. In connection withthis therapy, the compositions and combinations described are suitablefor preventing cranial neurologic damage and preventing and treatingcervicogenic headache.

[0071] The disclosed IL-18 antagonists, compositions and combinationtherapies are further used to treat conditions of the liver. For examplesoluble IL-18 receptor, alone or in combination with a TNF inhibitorsuch as ENBREL or other active agents, can be used to treat hepatitis,including acute alcoholic hepatitis, acute drug-induced or viralhepatitis, hepatitis A, B and C, sclerosing cholangitis and inflammationof the liver due to unknown causes. The invention is particularly usefulin treating hepatitis due to Hepatitis C virus. In connection with liverinflammation, IL-18 antagonists are further useful in treating hepaticsinusoid epithelium and biliary atresia.

[0072] In addition, the disclosed IL-18 antagonists, compositions andcombination therapies are used to treat various disorders that involvehearing loss and that are associated with abnormal IL-18 expression. Forexample, soluble IL-18 receptor, alone or in combination with TNFinhibitors, can be used to treat or prevent cochlear nerve-associatedhearing loss that is thought to result from an autoimmune process, i.e.,autoimmune hearing loss. This condition currently is treated withsteroids, methotrexate and/or cyclophosphamide. Also treatable orpreventable with the disclosed IL-18 antagonists, compositions andcombination therapies is Meniere's syndrome and Scholesteatoma, a middleear disorder often associated with hearing loss.

[0073] In addition, the subject invention provides IL-18 antagonists,e.g. soluble IL-18 receptor or IL-18 binding protein or an antagonisticantibody, compositions and combination therapies (e.g. soluble IL-18receptor and a TNF inhibitor such as ENBREL or other active agents) forthe treatment of non-arthritic medical conditions of the bones andjoints. This encompasses osteoarthritis and periodontitis resulting intooth loosening or loss, and prosthesis loosening after jointreplacement (generally associated with an inflammatory response to weardebris). This latter condition also is called “orthopedic implantosteolysis.” Another condition treatable with the compounds,compositions and combination therapies of the invention is temporalmandibular joint dysfunction (TMJ).

[0074] The following pulmonary disorders also can be treated orprevented with the disclosed IL-18 antagonists, compositions andcombination therapies (e.g. IL-18 antagonist and a TNF inhibitor such asENBREL or other active agents): adult respiratory distress syndrome(ARDS), acute respiratory distress syndrome and acute lung injury causedby a variety of conditions, including exposure to toxic chemicals,pancreatitis, trauma or other causes of inflammation. The disclosedcompounds, compositions and combination therapies of the invention alsoare useful for treating broncho-pulmonary dysplasia (BPD); chronicobstructive pulmonary diseases (e.g. emphysema and chronic bronchitis),and chronic fibrotic lung disease of preterm infants. In addition, thecompounds, compositions and combination therapies of the invention areused to treat occupational lung diseases, including asbestosis, coalworker's pneumoconiosis, silicosis or similar conditions associated withlong-term exposure to fine particles. hi other aspects of the invention,the disclosed compounds, compositions and combination therapies are usedto treat pulmonary fibrosis, including idiopathic pulmonary fibrosis andradiation-induced pulmonary fibrosis; pulmonary sarcoidosis; andallergies, including allergic rhinitis, contact dermatitis, atopicdermatitis and asthma.

[0075] The IL-18 antagonists, e.g. soluble IL-18 receptor, compositionsand combination therapies (e.g. an IL-18 antagonist as soluble IL-18receptor in combination with ENBREL or other TNF inhibitor or activeagent) of the invention are useful for treating or preventing primaryamyloidosis. In addition, the secondary amyloidosis that ischaracteristic of various conditions also are treatable with IL-18antagonists such as soluble IL-18 receptor, and the compositions andcombination therapies described herein. Such conditions include:Alzheimer's disease, secondary reactive amyloidosis; Down's syndrome;and dialysis-associated amyloidosis. Also treatable with the compounds,compositions and combination therapies of the invention are inheritedperiodic fever syndromes, including familial Mediterranean fever,hyperimmunoglobulin D and periodic fever syndrome and TNF-receptorassociated periodic syndromes (TRAPS).

[0076] Disorders involving the skin or mucous membranes also aretreatable using the disclosed IL-18 antagonists, compositions orcombination therapies, e.g. soluble IL-18 receptor and ENBREL. Suchdisorders include acantholytic diseases, including Darier's disease,keratosis follicularis and pemphigus vulgaris. Also treatable with thesubject IL-18 antagonists, especially soluble IL-18 receptor,compositions and combination therapies are acne; acne rosacea; alopeciaareata; aphthous stomatitis; bullous pemphigoid; burns; eczema;erythema, including erythema multiforme and erythema multiforme bullosum(Stevens-Johnson syndrome); inflammatory skin disease; lichen planus;linear IgA bullous disease (chronic bullous dermatosis of childhood);loss of skin elasticity; mucosal surface ulcers; neutrophilic dermatitis(Sweet's syndrome); dermatomyositis, pityriasis rubra pilaris;psoriasis; pyoderma gangrenosum; multicentric reticulohistiocytosis; andtoxic epidermal necrolysis.

[0077] Disorders associated with transplantation also are treatable orpreventable with the disclosed IL-18 antagonists, such as soluble IL-18receptor, compositions or combination therapies, including compositionsof soluble IL-18 receptor and ENBREL. Such disorders includegraft-versus-host disease, and complications resulting from solid organtransplantation, such as heart, liver, skin, kidney, lung (lungtransplant airway obliteration) or other transplants.

[0078] Ocular disorders also are treatable or preventable with thedisclosed IL-18 antagonists, especially soluble IL-18 receptor,compositions or combination therapies, including rhegmatogenous retinaldetachment, and inflammatory eye disease, including inflammatory eyedisease associated with smoking and macular degeneration.

[0079] IL-18 antagonists such as soluble IL-18 receptor and thedisclosed compositions and combination therapies also are useful fortreating disorders that affect the female reproductive system. Examplesinclude, but are not limited to, multiple implant failure/infertility;fetal loss syndrome or IV embryo loss (spontaneous abortion);preeclamptic pregnancies or eclampsia; endometriosis, chroniccervicitis, and pre-term labor.

[0080] In addition, the disclosed IL-18 antagonists, particularlysoluble IL-18 receptor or IL-18 binding protein or an antagonisticantibody, compositions and combination therapies, such as combinationsof IL-18 antagonist and ENBREL are useful for treating obesity,including to bring about a decrease in leptin formation. Also, thecompounds, compositions and combination therapies of the invention areused to treat or prevent sciatica, symptoms of aging, severe drugreactions (for example, IL-2 toxicity or bleomycin-induced pneumopathyand fibrosis), or to suppress the inflammatory response prior, during orafter the transfusion of allogeneic red blood cells in cardiac or othersurgery, or in treating a traumatic injury to a limb or joint, such astraumatic knee injury. Various other medical disorders treatable withthe disclosed IL-18 antagonists, compositions and combination therapiesinclude; multiple sclerosis; Behcet's syndrome; Sjogren's syndrome;autoimmune hemolytic anemia; beta thalassemia; amyotrophic lateralsclerosis (Lou Gehrig's Disease); Parkinson's disease; and tenosynovitisof unknown cause, as well as various autoimmune disorders or diseasesassociated with hereditary deficiencies, including x-linked mentalretardation.

[0081] The disclosed IL-18 antagonists, particularly soluble IL-18receptor, compositions and combination therapies, e.g. soluble IL-18receptor and ENBREL, are useful for treating the effects of neurotoxicneurotransmitters discharged during excitation of inflammation in thecentral nervous system and to inhibit or prevent the development ofglial scars at sites of central nervous system injury. In connectionwith central nervous system medical conditions, IL-18 antagonists, aloneor in combination with TNF inhibitors and particularly IL-18 antagonistand/or ENBREL are useful in treating temporal lobe epilepsy.Furthermore, the disclosed IL-18 antagonists, particularly soluble IL-18receptor or soluble IL-18 binding protein or an antagonistic antibody,compositions and combination therapies, e.g. soluble IL-18 receptor andENBREL, furthermore are useful for treating acute polyneuropathy;anorexia nervosa; Bell's palsy; chronic fatigue syndrome; transmissibledementia, including Creutzfeld-Jacob disease; demyelinating neuropathy;Guillain-Barre syndrome; vertebral disc disease; Gulf war syndrome;chronic inflammatory demyelinating polyneuropathy, myasthenia gravis;silent cerebral ischemia; sleep disorders, including narcolepsy andsleep apnea; chronic neuronal degeneration; and stroke, includingcerebral ischemic diseases. Other diseases and medical conditions thatcan be treated or prevented by administering an IL-18 antagonist, suchas soluble IL-18 receptor, alone or in combination with a hereindescribed active agents, particularly a TNF inhibitor such as ENBREL,include anorexia and/or anorexic conditions, peritonitis, endotoxemiaand septic shock, granuloma formation, heat stroke, Churg-Strausssyndrome, chronic inflammation following acute infections such astuberculosis and leprosy, systemic sclerosis and hypertrophic scarring.In addition to IL-18 antagonists in combination with TNF inhibitors,IFN-alpha beta or gamma and/or IL-4 inhibitors are suitable for treatinghypertrophic scarring.

[0082] Provided herein are methods of treating or preventing psoriaticlesions that involve administering to a human patient a therapeuticallyeffective amount of an IL-18 antagonist. A preferred antagonist for thispurpose is a soluble antagonist such as a soluble IL-18 receptor orIL-18 binding protein or an antagonistic antibody to IL-18 or acomponent of the IL-18 receptor. The treatment is effective againstpsoriatic lesions that occur in patients who have ordinary psoriasis orpsoriatic arthritis. In addition, any of the combination therapiesenumerated above are useful for the treatment of psoriasis.

[0083] Patients are defined as having ordinary psoriasis if they lackthe more serious symptoms of psoriatic arthritis (e.g., distalinterphalangeal joint DIP involvement, enthesopathy, spondylitis anddactylitis), but exhibit one of the following: 1) inflamed swollen skinlesions covered with silvery white scale (plaque psoriasis or psoriasisvulgaris); 2) small red dots appearing on the trunk, arms or legs(guttate psoriasis); 3) smooth inflamed lesions without scaling in theflexural surfaces of the skin (inverse psoriasis); 4) widespreadreddening and exfoliation of fine scales, with or without itching andswelling (erythrodermic psoriasis); 5) blister-like lesions (pustularpsoriasis); 6) elevated inflamed scalp lesions covered by silvery whitescales (scalp psoriasis); 7) pitted fingernails, with or withoutyellowish discoloration, crumbling nails, or inflammation and detachmentof the nail from the nail bed (nail psoriasis).

[0084] In treating ordinary psoriasis, an IL-18 antagonist isadministered in an amount and for a time sufficient to induce animprovement in the patient's condition as measured according to anyindicator that reflects the severity of the patient's psoriatic lesions.One or more such indicators may be assessed for determining whether theamount of IL-18 antagonist and duration of treatment is sufficient. Inone preferred embodiment of the invention, the soluble IL-18 receptor isadministered in an amount and for a time sufficient to induce animprovement over baseline in either the psoriasis area and severityindex (PASI) or the Target Lesion Assessment Score. In anotherembodiment, both indicators are used. When PASI score is used as theindicator, treatment is regarded as sufficient when the patient exhibitsan at least 50% improvement in his or her PASI score, or alternatively,when the patient exhibits an at least 75% improvement in PASI score.Using the Psoriasis Target Lesion Assessment Score to measuresufficiency of treatment involves determining for an individualpsoriatic lesion whether improvement has occurred in one or more of thefollowing, each of which is separately scored: plaque elevation; amountand degree of scaling or degree of erythema; and target lesion responseto treatment. Psoriasis Target Lesion Assessment Score is determined byadding together the separate scores for all four of the aforementionedindicia, and determining the extent of improvement by comparing thebaseline score the score after treatment has been administered.

[0085] IL-18 antagonists such as an IL-18 receptor or IL-18 bindingprotein also can be administered in combination with GM-CSF, IL-2 andinhibitors of protein kinase A type 1 to enhance T cell proliferation inHIV-infected patients who are receiving anti-retroviral therapy.

[0086] Although administration of IL-18 has been described as useful infighting infections, many complications from infection arise as a resultof an overactive or insufficiently controlled immune response. Thus, thedisclosed IL-18 antagonists, compositions and combination therapiesdescribed herein are useful in medicines for treating bacterial, viralor protozoal infections, and complications resulting therefrom. One suchdisease is Mycoplasma pneumonia. In addition, provided herein is the useof soluble IL-18 antagonist compositions or combinations, particularlyin combination with ENBREL to treat AIDS and related conditions, such asAIDS dementia complex, AIDS associated wasting, lipidistrophy due toantiretroviral therapy; CMV (cytomegalovirus) and Kaposi's sarcoma.Furthermore provided herein is the use of soluble IL-18 antagonistcompositions or combinations for treating protozoal diseases, includingmalaria and schistosomiasis. Additionally provided is the use of anIL-18 antagonist to treat erythema nodosum leprosum; bacterial or viralmeningitis; tuberculosis, including pulmonary tuberculosis; andpneumonitis secondary to a bacterial or viral infection. Provided alsoherein is the use of IL-18 antagonist compositions or combinations toprepare medicaments for treating louse-borne relapsing fevers, such asthat caused by Borrelia recurrentis. IL-18 antagonist can also be usedto prepare a medicament for treating conditions caused by Herpesviruses, such as herpetic stromal keratitis, corneal lesions; andvirus-induced corneal disorders. In addition, IL-18 antagonistcompositions and combinations can be used in treating humanpapillomavirus infections. IL-18 antagonist is used also to preparemedicaments to treat influenza infection. Further, IL-18 antagonistcompositions and combinations can be used to treat sepsis due tomicrobial infection.

[0087] The invention having been described, the following examples areoffered by way of illustration, and not limitation.

EXAMPLE

[0088] Effect Of Antagonizing IL-18 In A Mouse Model Of RheumatoidArthritis

[0089] This experiment was designed to test effect of antagonizing IL-18in a mouse model of rheumatoid arthritis, the well-knowncollagen-induced arthritis model. As an IL-18 antagonist, a fusionprotein was made between the IL-18 binding protein and an Fc mutein. Theamino acid sequence of the resulting protein, IL-18BP-Fc, is given inSEQ ID NO: 5. This protein was transiently expressed in CV-1/EBNA cellsfollowing DEAE-Dextran transfection of an expression vector, andpurified from the culture supernatant on a protein A column. Purity wasassessed by PAGE at greater than 98%.

[0090] In each experiment, male DBA/1 mice were immunized with collagenon day -21 and were boosted on day 0. Mice were treated daily from days0-14 with EP injections of antagonists or control proteins. Theincidence and severity of arthritis was monitored in a blind fashion.Each paw is assigned a score from 0 to 4 as follows: 0=normal;1=swelling in 1 to 3 digits; 2=mild swelling in ankles, forepaws, ormore than 3 digits; 3=moderate swelling in multiple joints; 4=severeswelling with loss of function. Each paw is totaled for a cumulativescore/mouse. Then, cumulative scores are totaled for mice in each groupfor a mean clinical score.

[0091] In the first & second experiments, mice were treated with 150μg/day of IL-18BP-Fc (n=10 in each experiment) and compared to micetreated with 150 μg/day of either entanercept (ENBREL®) (huTNFRFc) or HuIgG (n=15/group in each experiment). In both experiments, mice treatedwith IL-18BP-Fc demonstrated a statistically significant reduction inthe incidence and severity of the disease (73% reduction in meanclinical score in the first experiment, and 88% reduction in meanclinical score in the second experiment) compared with controls. ENBREL®treatment was also very effective at inhibiting the disease (92%reduction in mean clinical score in the first experiment, and 90%reduction in mean clinical score in the second experiment) compared withcontrols.

EXAMPLE

[0092] Dose Response Experiment In A Mouse Model of Rheumatoid Arthritis

[0093] A third experiment in the same mouse model of CIA tested a doseresponse in which IL-18BP-FC was administered IP at 150, 50, 15, and 5μg/day. As in the previous experiment, male DBA/1 mice were immunizedwith collagen on day-21 and were boosted on day 0. Mice were treateddaily from days 0-14 with IP injections of antagonists or controlproteins. The incidence and severity of arthritis was monitored in ablind fashion.

[0094] Groups of 15 mice were treated with the indicated doses ofIL-18BP-Fc or with 150 μg/day of ENBREL® or Hu IgG. All doses ofIL-18BP-Fc tested significantly reduced the disease incidence andseverity of arthritis (63%-76% reduction in mean clinical score forIL-18BP-Fc treated mice; 85% reduction in mean clinical score for thehuTNFRFc treated group) compared with the control group.

[0095] The results indicate that at each dose level, IL-18BP-Fceffectively inhibited onset of CIA when the reagent is administered in apreventative protocol. Thus, the minimally effective dose of IL-18BP-Fcin the preventative protocol could not be deduced using this range ofdosages; an additional titration experiment is therefore done todetermine the minimally effective dosage.

EXAMPLE

[0096] Combination Treatment Of An IL-18 Antagonist with Entanercept(ENBREL®)

[0097] The minimally effective dose of an IL-18 antagonist, in thiscase, IL-18BP-Fc, is co-administered along with minimally effectivedoses of entanercept (ENBREL®) to mice with CIA. Induction of arthritisand controls are as described above. The combination of IL-18BP-Fc andentanercept (ENBREL®) effectively inhibits CIA.

EXAMPLE

[0098] Effect of Antagonizing IL-18 In A Therapeutic Mouse Model ofRheumatoid Arthritis

[0099] This experiment was designed to assess the effect of inhibitingIL-18 as a therapeutic agent after arthritis had been established. Thus,the protocol for initiation of arthritis via injections of collagen wasthe same as for the above experiments, except that treatment was notbegun until after the mice had established disease symptoms. In thisexperiment, this point occurred at day 7 after the boost (28 days afterthe first treatment with collagen). Treatment with the followingpolypeptides at the indicated dose (daily injections, ip) was thenperformed for 13 days. Each treatment group consisted of 8 miceexhibiting signs of the disease. As before, an irrelevant human IgG wasused as a negative control, and entanercept (ENBREL®) was used as apositive control. In addition, a monoclonal antibody to the IL-1R (M147)was also used as a positive control (Rogers et al., 1992, Proc. Natl.Acad. Sci. USA 89:1011-1015). TABLE 1 Treatment Dose Score after 13 daystreatment IL-18BP-Fc 300 ug 6.1 HuIgG 300 ug 10.1 Enbrel 150 ug 6.7anti-IL1R Ab (M147)  50 ug 0

[0100] These results demonstrated a beneficial effect of using an IL-18antagonist as a therapeutic agent after arthritis had been established.Administration of IL-18BP-Fc did prevent progression of diseasesymptoms, in a manner similar to the results seen using ENBREL® in thisanimal model system.

EXAMPLE

[0101] Antagonism of IL-18 In Two Different Mouse Models of InflammatoryBowel Disease (IBD) With p13Fc

[0102] This experiment was designed to determine whether IL-18 plays asignificant role in the pathology of IBD and, if so, can its effect beblocked in vivo with an IL-18 antagonist. The effect of blocking IL-18was analyzed in both an experimentally induced model of IBD (the DSSmodel), and a spontaneous mouse model of IBD.

[0103] One of the most widely used models of IBD is the DSS model(dextran sulphate salt). In this model, dextran sulphate salt (m.w. canbe 40,000 to 500,000, but usually use that around 40,000) is given tomice (or other small mammals) in their drinking water at 2% to 8%.

[0104] For the spontaneous mouse model of IBD, Mdr1 a knockout mice wereused. These mice, which are homozygous for a deletion of function in theMdr1 a locus, spontaneously develop inflammatory bowel disease.

[0105] In this experiment, p13Fc was used as the IL-18 antagonist. p13Fcis an Fc fusion derivative of a fowlpox viral protein that binds toIL-18 (Born et al., 2000, J. Immunol. 164(6):3246-54).

[0106] Mice and Experimental Design

[0107] Two experimental groups of mice were studied: C57B1/6 mice withDSS-induced colitis, and Mdrla knockout mice (Mdr1 a−/−) that developspontaneous colitis. An irrelevant human IgG (HuIg) was used as anegative control, and a monoclonal antibody M147 previously shown todecrease weight loss in DSS-induced IBD was used as a positive control.The experimental groups were set up as follows:

[0108] C57BL/6 mice; n=4 mice per group

[0109] No DSS+HuIg (150 μg/day/mouse D0-D8)

[0110] 3% DSS+p13Fc (150 μg/day/mouse D0-D8)

[0111] 3% DSS+HuIgG (150 ag/day/mouse D0-D8)

[0112] 3% DSS+M147 (250 μg/day/mouse D0-D8)

[0113] FVB and Mdr1a−/− mice; n=4 per group

[0114] Healthy FVB mice+HuIgG (150 μg/day/mouse D0-D10)

[0115] Sick Mdr1a−/− mice+p13Fc (150 μg/day/mouse D0-D10)

[0116] Sick Mdr1a−/− mice+HuIgG (150 μg/day/mouse D0-D10)

[0117] Sick Mdr1a−/− mice+M147 mAb (250 μg/day/mouse D0-D10)

[0118] Mice were weighed daily; weight loss is a clinical sign of thedisease. Tissues were harvested at day 8 (D8) in the DSS modelexperiment, and at day 19 (D10) in the Mdr1a−/− model experiment.Histopathology (2 mice) was performed on the following tissues: smallintestine, large intestine and mesenteric lymph nodes (MLN).

[0119] Results and Conclusion

[0120] Preliminary histological analysis indicates that in the DSSmodel, inflammation in the large intestine was mildly reduced. Theresults from these preliminary experiments demonstrated that p13Fc canattenuate some of the weight loss induced by DSS-induced colitis, andsome of the weight loss associated with ongoing exacerbation ofspontaneous colitis.

EXAMPLE

[0121] Antagonism of IL-18 In the DSS Mouse Model of Inflammatory BowelDisease (IBD) With IL-18BP

[0122] This experiment was designed to analyze the effect of blockingIL-18 with IL-18BP-Fc, another IL-18 antagonist, during experimentallyinduced IBD. The DSS model of IBD was used. Controls were as describedin the previous experiment. The sequence of the IL-18BP-Fc protein usedis given in SEQ ID NO: 5.

[0123] Mice and Experimental Design.

[0124] C57BL/6 mice were given 2% DSS from day 0 to day 7 (D0-7); n=4mice per group

[0125] Groups:

[0126] No DSS+huIgG (250 μg/day/mouse D0-D7)

[0127] 2% DSS+IL-18BP-Fc (60 μg/day/mouse D0-D7)

[0128] 2% DSS+IL-18BP-Fc (100 μg/day/mouse D0-D7)

[0129] 2% DSS+huIgG (250 μg/day/mouse D0-D7)

[0130] 2% DSS+M147 (250 μg/day/mouse D0-D7)

[0131] Mice were weighed daily. Tissues (intestine and MLN) wereharvested at day 8 (D8). Histopathology (2 mice) analyses were alsoperformed on the tissues.

[0132] Results and Conclusion

[0133] Preliminary histological analysis indicated that inflammation inthe large intestine was mildly reduced. The results from theseexperiments also showed that IL-18BP-Fc is able to attenuate some of theweight loss associated with DSS-induced colitis.

EXAMPLE

[0134] Dose Response Experiments Using IL-18BP-Fc In the DSS Mouse Modelof Inflammatory Bowel Disease (IBD)

[0135] In order to further examine the effect of antagonizing IL-18 in amouse model of IBD, IL-18BP-Fc was administered at a higher range ofdosages.

[0136] Mice and Experimental Design

[0137] C57BL/6 mice were given 2% DSS in their drinking water for 7 days(D0-7). Treatment groups were as below; each treatment group contained 8mice.

[0138] No DSS+huIgG (150 μg/day/mouse)

[0139] DSS+IL-18BP-Fc (300 μg/day/mouse)

[0140] DSS+IL-18BP-Fc (100 μg/day/mouse)

[0141] DSS +hulgG (150 μg/day/mouse)

[0142] DSS +M147 (250 μg/day/mouse)

[0143] Mice were analyzed for weight loss daily. At the end of theexperiment, large intestine, small intestine and MLN were taken forhistology and for RNA analysis by RNase protection assay. MLN cells werealso counted, and analyzed by flow cytometry (FACs) for cell phenotypingand for cytokine production in vitro by ELISA (after stimulation withanti-CD3).

[0144] Results and Conclusions

[0145] The higher dose of IL-18BP-Fc (300 μg) inhibited weight loss byabout 32-35% on Day 8 of treatment. IL-18BP-Fc also inhibited theincreased cellularity (increased number of total cells per MLN) that istypically seen in the MLN after DSS treatment, suggesting that it wasblocking the process of cellular infiltration during inflammation.

[0146] Cytokine production by MLN cells after stimulation with CD3 wasexamined. This analysis indicated that IL-18BP-Fc inhibited theincreased IFNy production that would otherwise occur during DSS-inducedcolitis. Histological data also showed decreased inflammation in thelarge intestine after administration of IL-18BP-Fc at the higher dose.

[0147] Using RNase Protection Analysis, it was also observed thatIL-18BP-Fc decreased the levels of RNA encoding IL-1α/β and the IL-1receptor antagonist (IL-1RA) in the DSS model. This result is indicativeof reduced IL-1 production and possibly reduced inflammation in the gut.

EXAMPLE

[0148] Effect of Increased Dosage Of IL-18BP-Fc In the DSS Mouse Modelof Inflammatory Bowel Disease (IBD)

[0149] One possibility for the moderate effect of IL-18BP-Fc that wasobserved in the previous experiment was that the dose was not optimal.Accordingly, another experiment was performed with a higher dose ofIL-18BP-Fc (600 μg/mouse/day).

[0150] Mice and Experimental Design #/ treatment Mice group 2% DSS(D-2--> D7) C57Bl/6 6 No None C57BL/6 6 Yes 250 μg/day HuIgG C57Bl/6 6Yes 600 μg/day IL-18BP-Fc C57BL/6 6 Yes 250 μg/day M147

[0151] Mice were analyzed for weight loss daily. At the end of theexperiment, large intestine, small intestine and MLN were taken forhistology and for RNA analysis. The number of cells in each MLN wascounted (by dilution and staining of a sample) and an average for eachgroup determined. In addition, the MLN cells were analyzed by flowcytometry (FACs) for cell phenotyping and for cytokine production invitro by ELISA (after stimulation with anti-CD3).

[0152] Results and Conclusions

[0153] The effect of treatment on weight loss over the 8 day course ofthe experiment is illustrated in FIG. 1. Both M147 and IL-18BP-Fcsignificantly inhibited weight loss (85-90%) in the DSS-induced colitismodel. Histological analysis indicated that there was reducedinflammation in the large intestine in the IL-18BP-Fc-treated andM147-treated groups, as compared to mice treated with a control antibody(human IgG).

[0154] DSS treatment increases the levels of MLN cellularity (averagenumber of cells per MLN per treatment group) by about 2-fold. MLNcellularity was decreased to control levels (that seen with no DSStreatment) in IL-18BP-Fc-treated and M147-treated mice. The MLN cellsfrom DSS-treated mice, when stimulated by CD3, drastically increase IFNyand IL-10 production as compared to control (no DSS) treated mice.Administration of either IL-18BP-Fc or M147 attenuated significantlythis response (see FIG. 2A, which illustrates the average level of IFNγproduction from each treatment group, and FIG. 2B, which illustrates theaverage level of IL-10 production).

[0155] RNA analysis using both RNase Protection Assays as well as DNAarrays showed reduced levels of the mRNAs encoding multiple differentindicators of inflammation. These indicators included inflammatorycytokines such as IL-1α/β, TNFα and IFNγ, as well as proteins involvedin tissue repair including matrix metalloproteinases (MMPs). The resultsof RNase Protection Assays (RPA) detecting mRNAs in the large intestineencoding for IFNγ, TNFα, IL-6, IL-10, IL-18 and IL-1RA are shown in FIG.3A. The results from an RNase Protection Assay (RPA) detecting mRNAs inthe large intestine encoding for IL-1α and IL-β are shown in FIG. 3B.These results show that upregulation of inflammatory genes by DSStreatment was attenuated when IL-18BP-Fc or M147 are administered. IL-18RNA was however not regulated after IL-18BP-Fc treatment indicating thattranscription of IL-18 was not affected in IL-18BP-Fc treated mice, andsuggesting that IL-18BP-Fc does not regulate IL-18 by a transcriptionalfeedback mechanism. In conclusion, administration of IL-18BP-Fc clearlyattenuated the weight loss and inflammation associated with DSS-inducedcolitis in mice.

EXAMPLE

[0156] Effect of IL-18BP-Fc In the DSS Mouse Model of Inflammatory BowelDisease (IBD) On MLN Cellularity and Chemokine Secretion

[0157] One of the major characteristics of inflammation in thegastrointestinal tract is that the cellularity of the MLN, the majorlymph node draining from the gut, increases compared with non-coliticmice. This change in cellularity is due to infiltrating mononuclearcells, such as T cells and macrophages. Analysis of T cell populationsand numbers in the MLN revealed an increase in the relative proportionand absolute number of CD8⁺T cells compared with CD4⁺T cells. IL-18BP-Fcinhibited the increased cellularity seen in MLN of mice with DSS-colitisand analysis of the T cell populations and CD4⁺/CD8⁺T cell ratios showedthat the numbers and the ratios of T cells in the MLN fromIL-18BP-Fc-treated mice are similar to non-DSS treated groups.

[0158] Because cytokines have been shown to play an important role inmultiple models of colitis and as we observed changes in cytokine mRNAprofiles in the intestine, we were interested in determining whethercytokine protein profiles of cells draining from the gut would bemodulated by IL-18bp treatment. We analyzed MLN from the various groupsof animals on d8 for the in vitro secretion of IL-4, IL-10 andIFN-alpha. MLN cells were cultured on either PBS or anti-CD3 coatedplates for 48 hours and the culture supernatants analyzed by ELISA forcytokines. MLN taken from mice with DSS colitis showed increased levelsof both IFN-alpha and IL-10 protein production following anti-CD3treatment. In contrast, MLN from IL-18BP-Fc treated DSS colitic animalsdid not show the same increase in the levels of IFN-alpha and IL-10protein. Although the protein levels were not identical to controlnon-DSS levels, the decrease for IFN-alpha was significant (p=0.003).IL-4 was below detectable levels for all samples in these experiments(data not shown). These results show that treatment with IL-18bpinhibits the process of inflammation during DSS-induced colitispresumably by attenuating the trafficking of T cells into the MLN andthus attenuating the increased cytokine secretion in the gut associatedlymphoid tissues.

EXAMPLE

[0159] Analysis of Chemokine/Chemokine Receptor and MMP Gene Regulationin IL-18BP-Fc Treated Mice During DSS Colitis

[0160] To further characterize an expanded set of genes, RNA from the LIof the animals from the various treatment groups (d8) were used in arrayanalysis using Affymetrix chips. Approximately 300 genes showed greaterthan 3-fold regulation after DSS treatment and counter regulationfollowing IL-18BP-Fc treatment.

[0161] Focussing on chemokine and chemokine receptor gene regulation, weobserved increases in MIP1-alpha, MIP1-beta, MIP2, RANTES, CCR2, andCCR5 during DSS colitis. Treatment with IL-18BP-Fc attenuated theupregulation of these genes indicating that IL-18bp treatment may actupstream and be able to block the initiation of the chemokineinflammatory cascade associated with colitis. We also saw upregulationof ENA-78 (24×) and MIG (20×) in the LI of DSS colitic mice. IL-18BP-Fctreatment downregulated expression of these genes. These data areconsistent with our histopathological analysis shown in Table I thatindicates less recruitment of cells to the mucosa in IL-18bp treatedmice.

[0162] With regards to tissue repair and remodeling mechanisms duringinflammation, a number of investigators have documented an increase inexpression of a number of MMPs in human IBD including stromelysins(MMP-3 and 10), gelatinase B (MMP-9), collagenases (MMP-1,8 and 13) andtype IV collagen, as well as increases in tissue inhibitor ofmetalloproteinase (TIMP-1). Here we report an increase in RNA levels forMMP-3, 7, 9, 10, 13 and TIMP-1 in the LI of mice treated with DSS.Treatment with IL-18BP-Fc decreases mRNA levels for these MMPs andTIMP-1, down to that seen in control tissue, again indicating thatblocking IL-18 attenuates the damage incurred during the initiatinginflammatory stages of IBD.

[0163] In summary, these examples clearly demonstrate that blockingIL-18 function in vivo is an effective method of attenuating intestinalinflammation induced by DSS. These data indicate a role for the IL-18pathway in the initiation of intestinal damage associated with IBD, thatthat IL-18 antagonists can be used as a therapeutic approach fortreating IBD in humans.

[0164] The present invention is not to be limited in scope by thespecific embodiments described herein, which are intended as singleillustrations of individual aspects of the invention, and functionallyequivalent methods and components are within the scope of the invention.Indeed, various modifications of the invention, in addition to thoseshown and described herein will become apparent to those skilled in theart from the foregoing description and accompanying drawings. Suchmodifications are intended to fall within the scope of the appendedclaims.

1 5 1 2681 DNA Homo sapiens CDS (484)..(2283) 1 ctctctggat aggaagaaatatagtagaac cctttgaaaa tggatatttt cacatatttt 60 cgttcagata caaaagctggcagttactga aataaggact tgaagttcct tcctcttttt 120 ttatgtctta agagcaggaaataaagagac agctgaaggt gtagccttga ccaactgaaa 180 gggaaatctt catcctctgaaaaaacatat gtgattctca aaaaacgcat ctggaaaatt 240 gataaagaag cgattctgtagattctccca gcgctgttgg gctctcaatt ccttctgtga 300 aggacaacat atggtgatggggaaatcaga agctttgaga ccctctacac ctggatatga 360 atcccccttc taatacttaccagaaatgaa ggggatactc agggcagagt tctgaatctc 420 aaaacactct actctggcaaaggaatgaag ttattggagt gatgacagga acacgggaga 480 aca atg ctc tgt ttg ggctgg ata ttt ctt tgg ctt gtt gca gga gag 528 Met Leu Cys Leu Gly Trp IlePhe Leu Trp Leu Val Ala Gly Glu 1 5 10 15 cga att aaa gga ttt aat atttca ggt tgt tcc aca aaa aaa ctc ctt 576 Arg Ile Lys Gly Phe Asn Ile SerGly Cys Ser Thr Lys Lys Leu Leu 20 25 30 tgg aca tat tct aca agg agt gaagag gaa ttt gtc tta ttt tgt gat 624 Trp Thr Tyr Ser Thr Arg Ser Glu GluGlu Phe Val Leu Phe Cys Asp 35 40 45 tta cca gag cca cag aaa tca cat ttctgc cac aga aat cga ctc tca 672 Leu Pro Glu Pro Gln Lys Ser His Phe CysHis Arg Asn Arg Leu Ser 50 55 60 cca aaa caa gtc cct gag cac ctg ccc ttcatg ggt agt aac gac cta 720 Pro Lys Gln Val Pro Glu His Leu Pro Phe MetGly Ser Asn Asp Leu 65 70 75 tct gat gtc caa tgg tac caa caa cct tcg aatgga gat cca tta gag 768 Ser Asp Val Gln Trp Tyr Gln Gln Pro Ser Asn GlyAsp Pro Leu Glu 80 85 90 95 gac att agg aaa agc tat cct cac atc att caggac aaa tgt acc ctt 816 Asp Ile Arg Lys Ser Tyr Pro His Ile Ile Gln AspLys Cys Thr Leu 100 105 110 cac ttt ttg acc cca ggg gtg aat aat tct gggtca tat att tgt aga 864 His Phe Leu Thr Pro Gly Val Asn Asn Ser Gly SerTyr Ile Cys Arg 115 120 125 ccc aag atg att aag agc ccc tat gat gta gcctgt tgt gtc aag atg 912 Pro Lys Met Ile Lys Ser Pro Tyr Asp Val Ala CysCys Val Lys Met 130 135 140 att tta gaa gtt aag ccc cag aca aat gca tcctgt gag tat tcc gca 960 Ile Leu Glu Val Lys Pro Gln Thr Asn Ala Ser CysGlu Tyr Ser Ala 145 150 155 tca cat aag caa gac cta ctt ctt ggg agc actggc tct att tct tgc 1008 Ser His Lys Gln Asp Leu Leu Leu Gly Ser Thr GlySer Ile Ser Cys 160 165 170 175 ccc agt ctc agc tgc caa agt gat gca caaagt cca gcg gta acc tgg 1056 Pro Ser Leu Ser Cys Gln Ser Asp Ala Gln SerPro Ala Val Thr Trp 180 185 190 tac aag aat gga aaa ctc ctc tct gtg gaaagg agc aac cga atc gta 1104 Tyr Lys Asn Gly Lys Leu Leu Ser Val Glu ArgSer Asn Arg Ile Val 195 200 205 gtg gat gaa gtt tat gac tat cac cag ggcaca tat gta tgt gat tac 1152 Val Asp Glu Val Tyr Asp Tyr His Gln Gly ThrTyr Val Cys Asp Tyr 210 215 220 act cag tcg gat act gtg agt tcg tgg acagtc aga gct gtt gtt caa 1200 Thr Gln Ser Asp Thr Val Ser Ser Trp Thr ValArg Ala Val Val Gln 225 230 235 gtg aga acc att gtg gga gac act aaa ctcaaa cca gat att ctg gat 1248 Val Arg Thr Ile Val Gly Asp Thr Lys Leu LysPro Asp Ile Leu Asp 240 245 250 255 cct gtc gag gac aca ctg gaa gta gaactt gga aag cct tta act att 1296 Pro Val Glu Asp Thr Leu Glu Val Glu LeuGly Lys Pro Leu Thr Ile 260 265 270 agc tgc aaa gca cga ttt ggc ttt gaaagg gtc ttt aac cct gtc ata 1344 Ser Cys Lys Ala Arg Phe Gly Phe Glu ArgVal Phe Asn Pro Val Ile 275 280 285 aaa tgg tac atc aaa gat tct gac ctagag tgg gaa gtc tca gta cct 1392 Lys Trp Tyr Ile Lys Asp Ser Asp Leu GluTrp Glu Val Ser Val Pro 290 295 300 gag gcg aaa agt att aaa tcc act ttaaag gat gaa atc att gag cgt 1440 Glu Ala Lys Ser Ile Lys Ser Thr Leu LysAsp Glu Ile Ile Glu Arg 305 310 315 aat atc atc ttg gaa aaa gtc act cagcgt gat ctt cgc agg aag ttt 1488 Asn Ile Ile Leu Glu Lys Val Thr Gln ArgAsp Leu Arg Arg Lys Phe 320 325 330 335 gtt tgc ttt gtc cag aac tcc attgga aac aca acc cag tcc gtc caa 1536 Val Cys Phe Val Gln Asn Ser Ile GlyAsn Thr Thr Gln Ser Val Gln 340 345 350 ctg aaa gaa aag aga gga gtg gtgctc ctg tac atc ctg ctt ggc acc 1584 Leu Lys Glu Lys Arg Gly Val Val LeuLeu Tyr Ile Leu Leu Gly Thr 355 360 365 atc ggg acc ctg gtg gcc gtg ctggcg gcg agt gcc ctc ctc tac agg 1632 Ile Gly Thr Leu Val Ala Val Leu AlaAla Ser Ala Leu Leu Tyr Arg 370 375 380 cac tgg att gaa ata gtg ctg ctgtac cgg acc tac cag agc aag gat 1680 His Trp Ile Glu Ile Val Leu Leu TyrArg Thr Tyr Gln Ser Lys Asp 385 390 395 cag acg ctt ggg gat aaa aag gatttt gat gct ttc gta tcc tat gca 1728 Gln Thr Leu Gly Asp Lys Lys Asp PheAsp Ala Phe Val Ser Tyr Ala 400 405 410 415 aaa tgg agc tct ttt cca agtgag gcc act tca tct ctg agt gaa gaa 1776 Lys Trp Ser Ser Phe Pro Ser GluAla Thr Ser Ser Leu Ser Glu Glu 420 425 430 cac ttg gcc ctg agc cta tttcct gat gtt tta gaa aac aaa tat gga 1824 His Leu Ala Leu Ser Leu Phe ProAsp Val Leu Glu Asn Lys Tyr Gly 435 440 445 tat agc ctg tgt ttg ctt gaaaga gat gtg gct cca gga gga gtg tat 1872 Tyr Ser Leu Cys Leu Leu Glu ArgAsp Val Ala Pro Gly Gly Val Tyr 450 455 460 gca gaa gac att gtg agc attatt aag aga agc aga aga gga ata ttt 1920 Ala Glu Asp Ile Val Ser Ile IleLys Arg Ser Arg Arg Gly Ile Phe 465 470 475 atc ttg agc ccc aac tat gtcaat gga ccc agt atc ttt gaa cta caa 1968 Ile Leu Ser Pro Asn Tyr Val AsnGly Pro Ser Ile Phe Glu Leu Gln 480 485 490 495 gca gca gtg aat ctt gccttg gat gat caa aca ctg aaa ctc att tta 2016 Ala Ala Val Asn Leu Ala LeuAsp Asp Gln Thr Leu Lys Leu Ile Leu 500 505 510 att aag ttc tgt tac ttccaa gag cca gag tct cta cct cat ctc gtg 2064 Ile Lys Phe Cys Tyr Phe GlnGlu Pro Glu Ser Leu Pro His Leu Val 515 520 525 aaa aaa gct ctc agg gttttg ccc aca gtt act tgg aga ggc tta aaa 2112 Lys Lys Ala Leu Arg Val LeuPro Thr Val Thr Trp Arg Gly Leu Lys 530 535 540 tca gtt cct ccc aat tctagg ttc tgg gcc aaa atg cgc tac cac atg 2160 Ser Val Pro Pro Asn Ser ArgPhe Trp Ala Lys Met Arg Tyr His Met 545 550 555 cct gtg aaa aac tct caggga ttc acg tgg aac cag ctc aga att acc 2208 Pro Val Lys Asn Ser Gln GlyPhe Thr Trp Asn Gln Leu Arg Ile Thr 560 565 570 575 tct agg att ttt cagtgg aaa gga ctc agt aga aca gaa acc act ggg 2256 Ser Arg Ile Phe Gln TrpLys Gly Leu Ser Arg Thr Glu Thr Thr Gly 580 585 590 agg agc tcc cag cctaag gaa tgg tga aatgagccct ggagccccct 2303 Arg Ser Ser Gln Pro Lys GluTrp 595 ccagtccagt ccctgggata gagatgttgc tggacagaac tcacagctctgtgtgtgtgt 2363 gttcaggctg ataggaaatt caaagagtct cctgccagca ccaagcaagcttgatggaca 2423 atggaatggg attgagactg tggtttagag cctttgattt cctggactggacagacggcg 2483 agtgaattct ctagaccttg ggtactttca gtacacaaca cccctaagatttcccagtgg 2543 tccgagcaga atcagaaaat acagctactt ctgccttatg gctagggaactgtcatgtct 2603 accatgtatt gtacatatga ctttatgtat acttgcaatc aaataaatattattttatta 2663 gaaaaaaaac cggaattc 2681 2 599 PRT Homo sapiens 2 MetLeu Cys Leu Gly Trp Ile Phe Leu Trp Leu Val Ala Gly Glu Arg 1 5 10 15Ile Lys Gly Phe Asn Ile Ser Gly Cys Ser Thr Lys Lys Leu Leu Trp 20 25 30Thr Tyr Ser Thr Arg Ser Glu Glu Glu Phe Val Leu Phe Cys Asp Leu 35 40 45Pro Glu Pro Gln Lys Ser His Phe Cys His Arg Asn Arg Leu Ser Pro 50 55 60Lys Gln Val Pro Glu His Leu Pro Phe Met Gly Ser Asn Asp Leu Ser 65 70 7580 Asp Val Gln Trp Tyr Gln Gln Pro Ser Asn Gly Asp Pro Leu Glu Asp 85 9095 Ile Arg Lys Ser Tyr Pro His Ile Ile Gln Asp Lys Cys Thr Leu His 100105 110 Phe Leu Thr Pro Gly Val Asn Asn Ser Gly Ser Tyr Ile Cys Arg Pro115 120 125 Lys Met Ile Lys Ser Pro Tyr Asp Val Ala Cys Cys Val Lys MetIle 130 135 140 Leu Glu Val Lys Pro Gln Thr Asn Ala Ser Cys Glu Tyr SerAla Ser 145 150 155 160 His Lys Gln Asp Leu Leu Leu Gly Ser Thr Gly SerIle Ser Cys Pro 165 170 175 Ser Leu Ser Cys Gln Ser Asp Ala Gln Ser ProAla Val Thr Trp Tyr 180 185 190 Lys Asn Gly Lys Leu Leu Ser Val Glu ArgSer Asn Arg Ile Val Val 195 200 205 Asp Glu Val Tyr Asp Tyr His Gln GlyThr Tyr Val Cys Asp Tyr Thr 210 215 220 Gln Ser Asp Thr Val Ser Ser TrpThr Val Arg Ala Val Val Gln Val 225 230 235 240 Arg Thr Ile Val Gly AspThr Lys Leu Lys Pro Asp Ile Leu Asp Pro 245 250 255 Val Glu Asp Thr LeuGlu Val Glu Leu Gly Lys Pro Leu Thr Ile Ser 260 265 270 Cys Lys Ala ArgPhe Gly Phe Glu Arg Val Phe Asn Pro Val Ile Lys 275 280 285 Trp Tyr IleLys Asp Ser Asp Leu Glu Trp Glu Val Ser Val Pro Glu 290 295 300 Ala LysSer Ile Lys Ser Thr Leu Lys Asp Glu Ile Ile Glu Arg Asn 305 310 315 320Ile Ile Leu Glu Lys Val Thr Gln Arg Asp Leu Arg Arg Lys Phe Val 325 330335 Cys Phe Val Gln Asn Ser Ile Gly Asn Thr Thr Gln Ser Val Gln Leu 340345 350 Lys Glu Lys Arg Gly Val Val Leu Leu Tyr Ile Leu Leu Gly Thr Ile355 360 365 Gly Thr Leu Val Ala Val Leu Ala Ala Ser Ala Leu Leu Tyr ArgHis 370 375 380 Trp Ile Glu Ile Val Leu Leu Tyr Arg Thr Tyr Gln Ser LysAsp Gln 385 390 395 400 Thr Leu Gly Asp Lys Lys Asp Phe Asp Ala Phe ValSer Tyr Ala Lys 405 410 415 Trp Ser Ser Phe Pro Ser Glu Ala Thr Ser SerLeu Ser Glu Glu His 420 425 430 Leu Ala Leu Ser Leu Phe Pro Asp Val LeuGlu Asn Lys Tyr Gly Tyr 435 440 445 Ser Leu Cys Leu Leu Glu Arg Asp ValAla Pro Gly Gly Val Tyr Ala 450 455 460 Glu Asp Ile Val Ser Ile Ile LysArg Ser Arg Arg Gly Ile Phe Ile 465 470 475 480 Leu Ser Pro Asn Tyr ValAsn Gly Pro Ser Ile Phe Glu Leu Gln Ala 485 490 495 Ala Val Asn Leu AlaLeu Asp Asp Gln Thr Leu Lys Leu Ile Leu Ile 500 505 510 Lys Phe Cys TyrPhe Gln Glu Pro Glu Ser Leu Pro His Leu Val Lys 515 520 525 Lys Ala LeuArg Val Leu Pro Thr Val Thr Trp Arg Gly Leu Lys Ser 530 535 540 Val ProPro Asn Ser Arg Phe Trp Ala Lys Met Arg Tyr His Met Pro 545 550 555 560Val Lys Asn Ser Gln Gly Phe Thr Trp Asn Gln Leu Arg Ile Thr Ser 565 570575 Arg Ile Phe Gln Trp Lys Gly Leu Ser Arg Thr Glu Thr Thr Gly Arg 580585 590 Ser Ser Gln Pro Lys Glu Trp 595 3 1626 DNA Homo sapiens CDS(1)..(1626) 3 atg aat tgt aga gaa tta ccc ttg acc ctt tgg gtg ctt atatct gta 48 Met Asn Cys Arg Glu Leu Pro Leu Thr Leu Trp Val Leu Ile SerVal 1 5 10 15 agc act gca gaa tct tgt act tca cgt ccc cac att act gtggtt gaa 96 Ser Thr Ala Glu Ser Cys Thr Ser Arg Pro His Ile Thr Val ValGlu 20 25 30 ggg gaa cct ttc tat ctg aaa cat tgc tcg tgt tca ctt gca catgag 144 Gly Glu Pro Phe Tyr Leu Lys His Cys Ser Cys Ser Leu Ala His Glu35 40 45 att gaa aca acc acc aaa agc tgg tac aaa agc agt gga tca cag gaa192 Ile Glu Thr Thr Thr Lys Ser Trp Tyr Lys Ser Ser Gly Ser Gln Glu 5055 60 cat gtg gag ctg aac cca agg agt tcc tcg aga att gct ttg cat gat240 His Val Glu Leu Asn Pro Arg Ser Ser Ser Arg Ile Ala Leu His Asp 6570 75 80 tgt gtt ttg gag ttt tgg cca gtt gag ttg aat gac aca gga tct tac288 Cys Val Leu Glu Phe Trp Pro Val Glu Leu Asn Asp Thr Gly Ser Tyr 8590 95 ttt ttc caa atg aaa aat tat act cag aaa tgg aaa tta aat gtc atc336 Phe Phe Gln Met Lys Asn Tyr Thr Gln Lys Trp Lys Leu Asn Val Ile 100105 110 aga aga aat aaa cac agc tgt ttc act gaa aga caa gta act agt aaa384 Arg Arg Asn Lys His Ser Cys Phe Thr Glu Arg Gln Val Thr Ser Lys 115120 125 att gtg gaa gtt aaa aaa ttt ttt cag ata acc tgt gaa aac agt tac432 Ile Val Glu Val Lys Lys Phe Phe Gln Ile Thr Cys Glu Asn Ser Tyr 130135 140 tat caa aca ctg gtc aac agc aca tca ttg tat aag aac tgt aaa aag480 Tyr Gln Thr Leu Val Asn Ser Thr Ser Leu Tyr Lys Asn Cys Lys Lys 145150 155 160 cta cta ctg gag aac aat aaa aac cca acg ata aag aag aac gccgag 528 Leu Leu Leu Glu Asn Asn Lys Asn Pro Thr Ile Lys Lys Asn Ala Glu165 170 175 ttt gaa gat cag ggg tat tac tcc tgc gtg cat ttc ctt cat cataat 576 Phe Glu Asp Gln Gly Tyr Tyr Ser Cys Val His Phe Leu His His Asn180 185 190 gga aaa cta ttt aat atc acc aaa acc ttc aat ata aca ata gtggaa 624 Gly Lys Leu Phe Asn Ile Thr Lys Thr Phe Asn Ile Thr Ile Val Glu195 200 205 gat cgc agt aat ata gtt ccg gtt ctt ctt gga cca aag ctt aaccat 672 Asp Arg Ser Asn Ile Val Pro Val Leu Leu Gly Pro Lys Leu Asn His210 215 220 gtt gca gtg gaa tta gga aaa aac gta agg ctc aac tgc tct gctttg 720 Val Ala Val Glu Leu Gly Lys Asn Val Arg Leu Asn Cys Ser Ala Leu225 230 235 240 ctg aat gaa gag gat gta att tat tgg atg ttt ggg gaa gaaaat gga 768 Leu Asn Glu Glu Asp Val Ile Tyr Trp Met Phe Gly Glu Glu AsnGly 245 250 255 tcg gat cct aat ata cat gaa gag aaa gaa atg aga att atgact cca 816 Ser Asp Pro Asn Ile His Glu Glu Lys Glu Met Arg Ile Met ThrPro 260 265 270 gaa ggc aaa tgg cat gct tca aaa gta ttg aga att gaa aatatt ggt 864 Glu Gly Lys Trp His Ala Ser Lys Val Leu Arg Ile Glu Asn IleGly 275 280 285 gaa agc aat cta aat gtt tta tat aat tgc act gtg gcc agcacg gga 912 Glu Ser Asn Leu Asn Val Leu Tyr Asn Cys Thr Val Ala Ser ThrGly 290 295 300 ggc aca gac acc aaa agc ttc atc ttg gtg aga aaa gca gacatg gct 960 Gly Thr Asp Thr Lys Ser Phe Ile Leu Val Arg Lys Ala Asp MetAla 305 310 315 320 gat atc cca ggc cac gtc ttc aca aga gga atg atc atagct gtt ttg 1008 Asp Ile Pro Gly His Val Phe Thr Arg Gly Met Ile Ile AlaVal Leu 325 330 335 atc ttg gtg gca gta gtg tgc cta gtg act gtg tgt gtcatt tat aga 1056 Ile Leu Val Ala Val Val Cys Leu Val Thr Val Cys Val IleTyr Arg 340 345 350 gtt gac ttg gtt cta ttt tat aga cat tta acg aga agagat gaa aca 1104 Val Asp Leu Val Leu Phe Tyr Arg His Leu Thr Arg Arg AspGlu Thr 355 360 365 tta aca gat gga aaa aca tat gat gct ttt gtg tct taccta aaa gaa 1152 Leu Thr Asp Gly Lys Thr Tyr Asp Ala Phe Val Ser Tyr LeuLys Glu 370 375 380 tgc cga cct gaa aat gga gag gag cac acc ttt gct gtggag att ttg 1200 Cys Arg Pro Glu Asn Gly Glu Glu His Thr Phe Ala Val GluIle Leu 385 390 395 400 ccc agg gtg ttg gag aaa cat ttt ggg tat aag ttatgc ata ttt gaa 1248 Pro Arg Val Leu Glu Lys His Phe Gly Tyr Lys Leu CysIle Phe Glu 405 410 415 agg gat gta gtg cct gga gga gct gtt gtt gat gaaatc cac tca ctg 1296 Arg Asp Val Val Pro Gly Gly Ala Val Val Asp Glu IleHis Ser Leu 420 425 430 ata gag aaa agc cga aga cta atc att gtc cta agtaaa agt tat atg 1344 Ile Glu Lys Ser Arg Arg Leu Ile Ile Val Leu Ser LysSer Tyr Met 435 440 445 tct aat gag gtc agg tat gaa ctt gaa agt gga ctccat gaa gca ttg 1392 Ser Asn Glu Val Arg Tyr Glu Leu Glu Ser Gly Leu HisGlu Ala Leu 450 455 460 gtg gaa aga aaa att aaa ata atc tta att gaa tttaca cct gtt act 1440 Val Glu Arg Lys Ile Lys Ile Ile Leu Ile Glu Phe ThrPro Val Thr 465 470 475 480 gac ttc aca ttc ttg ccc caa tca cta aag cttttg aaa tct cac aga 1488 Asp Phe Thr Phe Leu Pro Gln Ser Leu Lys Leu LeuLys Ser His Arg 485 490 495 gtt ctg aag tgg aag gcc gat aaa tct ctt tcttat aac tca agg ttc 1536 Val Leu Lys Trp Lys Ala Asp Lys Ser Leu Ser TyrAsn Ser Arg Phe 500 505 510 tgg aag aac ctt ctt tac tta atg cct gca aaaaca gtc aag cca ggt 1584 Trp Lys Asn Leu Leu Tyr Leu Met Pro Ala Lys ThrVal Lys Pro Gly 515 520 525 aga gac gaa ccg gaa gtc ttg cct gtt ctt tccgag tct taa 1626 Arg Asp Glu Pro Glu Val Leu Pro Val Leu Ser Glu Ser 530535 540 4 541 PRT Homo sapiens 4 Met Asn Cys Arg Glu Leu Pro Leu Thr LeuTrp Val Leu Ile Ser Val 1 5 10 15 Ser Thr Ala Glu Ser Cys Thr Ser ArgPro His Ile Thr Val Val Glu 20 25 30 Gly Glu Pro Phe Tyr Leu Lys His CysSer Cys Ser Leu Ala His Glu 35 40 45 Ile Glu Thr Thr Thr Lys Ser Trp TyrLys Ser Ser Gly Ser Gln Glu 50 55 60 His Val Glu Leu Asn Pro Arg Ser SerSer Arg Ile Ala Leu His Asp 65 70 75 80 Cys Val Leu Glu Phe Trp Pro ValGlu Leu Asn Asp Thr Gly Ser Tyr 85 90 95 Phe Phe Gln Met Lys Asn Tyr ThrGln Lys Trp Lys Leu Asn Val Ile 100 105 110 Arg Arg Asn Lys His Ser CysPhe Thr Glu Arg Gln Val Thr Ser Lys 115 120 125 Ile Val Glu Val Lys LysPhe Phe Gln Ile Thr Cys Glu Asn Ser Tyr 130 135 140 Tyr Gln Thr Leu ValAsn Ser Thr Ser Leu Tyr Lys Asn Cys Lys Lys 145 150 155 160 Leu Leu LeuGlu Asn Asn Lys Asn Pro Thr Ile Lys Lys Asn Ala Glu 165 170 175 Phe GluAsp Gln Gly Tyr Tyr Ser Cys Val His Phe Leu His His Asn 180 185 190 GlyLys Leu Phe Asn Ile Thr Lys Thr Phe Asn Ile Thr Ile Val Glu 195 200 205Asp Arg Ser Asn Ile Val Pro Val Leu Leu Gly Pro Lys Leu Asn His 210 215220 Val Ala Val Glu Leu Gly Lys Asn Val Arg Leu Asn Cys Ser Ala Leu 225230 235 240 Leu Asn Glu Glu Asp Val Ile Tyr Trp Met Phe Gly Glu Glu AsnGly 245 250 255 Ser Asp Pro Asn Ile His Glu Glu Lys Glu Met Arg Ile MetThr Pro 260 265 270 Glu Gly Lys Trp His Ala Ser Lys Val Leu Arg Ile GluAsn Ile Gly 275 280 285 Glu Ser Asn Leu Asn Val Leu Tyr Asn Cys Thr ValAla Ser Thr Gly 290 295 300 Gly Thr Asp Thr Lys Ser Phe Ile Leu Val ArgLys Ala Asp Met Ala 305 310 315 320 Asp Ile Pro Gly His Val Phe Thr ArgGly Met Ile Ile Ala Val Leu 325 330 335 Ile Leu Val Ala Val Val Cys LeuVal Thr Val Cys Val Ile Tyr Arg 340 345 350 Val Asp Leu Val Leu Phe TyrArg His Leu Thr Arg Arg Asp Glu Thr 355 360 365 Leu Thr Asp Gly Lys ThrTyr Asp Ala Phe Val Ser Tyr Leu Lys Glu 370 375 380 Cys Arg Pro Glu AsnGly Glu Glu His Thr Phe Ala Val Glu Ile Leu 385 390 395 400 Pro Arg ValLeu Glu Lys His Phe Gly Tyr Lys Leu Cys Ile Phe Glu 405 410 415 Arg AspVal Val Pro Gly Gly Ala Val Val Asp Glu Ile His Ser Leu 420 425 430 IleGlu Lys Ser Arg Arg Leu Ile Ile Val Leu Ser Lys Ser Tyr Met 435 440 445Ser Asn Glu Val Arg Tyr Glu Leu Glu Ser Gly Leu His Glu Ala Leu 450 455460 Val Glu Arg Lys Ile Lys Ile Ile Leu Ile Glu Phe Thr Pro Val Thr 465470 475 480 Asp Phe Thr Phe Leu Pro Gln Ser Leu Lys Leu Leu Lys Ser HisArg 485 490 495 Val Leu Lys Trp Lys Ala Asp Lys Ser Leu Ser Tyr Asn SerArg Phe 500 505 510 Trp Lys Asn Leu Leu Tyr Leu Met Pro Ala Lys Thr ValLys Pro Gly 515 520 525 Arg Asp Glu Pro Glu Val Leu Pro Val Leu Ser GluSer 530 535 540 5 422 PRT Artificial Sequence IL-18 BP-Fc 5 Met Arg HisAsn Trp Thr Pro Asp Leu Ser Pro Leu Trp Val Leu Leu 1 5 10 15 Leu CysAla His Val Val Thr Leu Leu Val Arg Ala Thr Pro Val Ser 20 25 30 Gln ThrThr Thr Ala Ala Thr Ala Ser Val Arg Ser Thr Lys Asp Pro 35 40 45 Cys ProSer Gln Pro Pro Val Phe Pro Ala Ala Lys Gln Cys Pro Ala 50 55 60 Leu GluVal Thr Trp Pro Glu Val Glu Val Pro Leu Asn Gly Thr Leu 65 70 75 80 SerLeu Ser Cys Val Ala Cys Ser Arg Phe Pro Asn Phe Ser Ile Leu 85 90 95 TyrTrp Leu Gly Asn Gly Ser Phe Ile Glu His Leu Pro Gly Arg Leu 100 105 110Trp Glu Gly Ser Thr Ser Arg Glu Arg Gly Ser Thr Gly Thr Gln Leu 115 120125 Cys Lys Ala Leu Val Leu Glu Gln Leu Thr Pro Ala Leu His Ser Thr 130135 140 Asn Phe Ser Cys Val Leu Val Asp Pro Glu Gln Val Val Gln Arg His145 150 155 160 Val Val Leu Ala Gln Leu Trp Ala Gly Leu Arg Ala Thr LeuPro Pro 165 170 175 Thr Gln Glu Ala Leu Pro Ser Ser His Ser Ser Pro GlnGln Gln Gly 180 185 190 Arg Ser Cys Asp Lys Thr His Thr Cys Pro Pro CysPro Ala Pro Glu 195 200 205 Ala Glu Gly Ala Pro Ser Val Phe Leu Phe ProPro Lys Pro Lys Asp 210 215 220 Thr Leu Met Ile Ser Arg Thr Pro Glu ValThr Cys Val Val Val Asp 225 230 235 240 Val Ser His Glu Asp Pro Glu ValLys Phe Asn Trp Tyr Val Asp Gly 245 250 255 Val Glu Val His Asn Ala LysThr Lys Pro Arg Glu Glu Gln Tyr Asn 260 265 270 Ser Thr Tyr Arg Val ValSer Val Leu Thr Val Leu His Gln Asp Trp 275 280 285 Leu Asn Gly Lys GluTyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 290 295 300 Ala Pro Ile GluLys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 305 310 315 320 Pro GlnVal Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 325 330 335 GlnVal Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 340 345 350Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 355 360365 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 370375 380 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys385 390 395 400 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln LysSer Leu 405 410 415 Ser Leu Ser Pro Gly Lys 420

What is claimed is:
 1. A method of treating a patient afflicted with amedical disorder selected from the group consisting of rheumatoidarthritis and inflammatory bowel disease, the method comprisingadministering to said patient a therapeutically effective amount of anIL-18 antagonist.
 2. The method of claim 1, wherein the IL-18 antagonistis soluble IL-18 receptor.
 3. The method of claim 2, wherein the solubleIL-18 receptor is a heterodimeric receptor.
 4. The method of claim 1,wherein the IL-18 antagonist is an antibody.
 5. The method of claim 4,wherein the antibody immunospecifically recognizes a component of anIL-18 receptor.
 6. The method of claim 4, wherein the antibody is ahumanized antibody.
 7. The method of claim 6, wherein the antibody is asingle-chain antibody.
 8. The method of claim 1, wherein the IL-18antagonist is a soluble IL-18 binding protein.
 9. The method of claim 1,wherein the IL-18 antagonist is administered one or more times per week.10. The method of claim 1, wherein the IL-18 antagonist is administeredby subcutaneous injection.
 11. The method of claim 1, wherein the IL-18antagonist is administered in combination with one or more compoundsselected from the group consisting of non-steroidal anti-inflammatorydrugs; analgesics; systemic steroids; antagonists of inflammatorycytokines; anti-inflammatory cytokines; antibodies against T cellsurface proteins; anthralin; vitamin D3 and its analogs; oral retinoids;salicylic acid; methotrexate; cyclosporine; hydroxyurea; andsulfasalazine.
 12. The method of claim 1, wherein the IL-18 antagonistis administered in combination with a TNF inhibitor.
 13. The method ofclaim 12 wherein the TNF inhibitor is TNFR:Fc.
 14. The method of claim1, wherein the IL-18 antagonist is administered in combination with anantagonist to a cytokine selected from the group consisting of IFNγ,TGFβ, IL-6 and IL-8.
 15. The method of claim 12 wherein the IL-18antagonist and TNF inhibitor are administered in combination with anantagonist to a cytokine selected from the group consisting of IFNγ,TGFβ, IL-6, IL-8, IL-12, and IL-15.